Genetically altered lysm receptors with altered agonist specificity and affinity

ABSTRACT

Aspects of the present disclosure relates to genetically altered LysM receptors. In particular, the present disclosure relates to a hydrophobic patch into the LysM2 domain which can increase affinity and/or selectivity for LCOs and by replacement of regions in the LysM1 domain with the corresponding regions of the LysM1 domain from a donor LysM receptor that can alter the affinity and/or selectivity for the oligosaccharide particularly for LCOs and can alter the specificity between LCO when using regions from a high affinity and specificity LCO LysM receptor such as a legume NFR1 receptor. The present disclosure also relates to genetically altering LysM receptors in plants to include a hydrophobic patch or alter the hydrophobic patch and to genetically altering LysM receptors in plants by replacement of regions in the LysM2 domain.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/718,282, filed Aug. 13, 2018, which is hereby incorporated byreference in its entirety.

SUBMISSION OF SEQUENCE LISTING AS ASCII TEXT FILE

The content of the following submission on ASCII text file isincorporated herein by reference in its entirety: a computer readableform (CRF) of the Sequence Listing (file name: 794542000440SEQLIST.txt,date recorded: Aug. 12, 2019, size: 298 KB).

TECHNICAL FIELD

The present disclosure relates to genetically altered LysM receptors. Inparticular, the present disclosure relates to a hydrophobic patch intothe LysM2 domain which can increase affinity and/or selectivity for LCOsand by replacement of regions in the LysM1 domain with the correspondingregions of the LysM1 domain from a donor LysM receptor that can alterthe affinity and/or selectivity for the oligosaccharide, particularlyfor LCOs, and can alter the specificity between LCOs when using regionsfrom a high affinity and specificity LCO LysM receptor such as a legumeNFR1 receptor. The present disclosure also relates to geneticallyaltering LysM receptors in plants to include a hydrophobic patch oralter the hydrophobic patch and to genetically altering LysM receptorsin plants by replacement of regions in the LysM2 domain.

BACKGROUND

Plants are exposed to a wide variety of microbes in their environment,both benign and pathogenic. To protect against the pathogenic microbes,plants have the ability to recognize specific molecular signals of themicrobes through an array of receptors and, depending upon the patternof the signals, can initiate an appropriate immune response. Themolecular signals are derived from secreted materials, cell-wallcomponents, and even cytosolic proteins of the microbes.Chitooligosaccharides (COs) are an important fungal molecular signalthat plants recognize through the chitin receptors CEBiP, CERK1, LYK5,and CERK6 (previously called LYSE) found on the plasma membrane. Thesereceptors are in the LysM class of receptors and recognize the size andthe acetylation of COs from fungi. Lipo-chitooligosaccharides (LCOs) areanother important molecular signal that can be found on both bacteriaand fungi that are recognized by other LysM receptors.

In addition to benign and pathogenic microbes, some microbes can bebeneficial to plants through association or symbiosis. Plants that enterinto symbiotic relationships with certain nitrogen fixing bacteria andfungi need to be able to recognize the specific bacterial or fungalspecies to initiate the symbiosis while still being able to activatetheir immune systems to respond to other bacteria and fungi. Oneimportant mechanism that allows plants to recognize these specificbacteria or fungi is through specialized LysM receptors that have highaffinity, high selectivity, and/or high specificity for the form of LCOsproduced by the specific bacteria or fungi while LCOs from otherbacteria and fungi are not recognized by these specialized LysMreceptors.

Experimental and computational approaches have been used to identify anumber of these specialized LysM receptors (also referred to as highaffinity and specificity LCO receptors). As these receptors are requiredfor recognizing symbiotic bacterial and fungal species, and forinitiating symbiosis, these receptors represent an important componentof any plant engineering strategy. Using these receptors, however, willnot be particularly straightforward; transferring a specialized LysMreceptor into a plant that does not currently have one may require codonoptimization, the identification of suitable promoters, the use oftargeting signals, and further engineering approaches needed to adaptexogenous sequences for optimal expression. Further, the number of thesereceptors that have been identified is currently limited.

Moreover, species that already have specialized LysM receptors, e.g.,legumes, cannot be easily engineered with new specialized LysMreceptors. Currently, legumes are limited to the specific bacterial orfungal species with which they form symbiotic associations. Whilelegumes may have the benefit of existing symbiotic associations, theiragricultural potential is limited. For example, legumes cannot currentlybe easily engineered to have different specificity for differentsymbiotic microbial species, which would allow legumes to better formassociations with the bacterial or fungal species in different soils.Moreover, legumes cannot be easily engineered to have improvedspecialized LysM receptors. Further, legumes cannot currently beengineered to have synergistic symbiotic requirements with other cropsgrown in rotation with them. Editing approaches are needed for both themodification of endogenous LysM receptors into specialized LysMreceptors able to perceive symbiotic bacterial and fungal species, andthe modification of specialized LysM receptors into specialized LysMreceptors with different specific recognition of symbiotic bacterial andfungal species.

BRIEF SUMMARY

In order to meet these needs, the present disclosure providescomplementary means of modifying LysM receptors by introduction of ahydrophobic patch into the LysM2 domain which can increase affinityand/or selectivity for LCOs, and by replacement of regions in the LysM1domain with the corresponding regions of the LysM1 domain from a donorLysM receptor that can alter the affinity and/or selectivity for theoligosaccharide, particularly for LCOs, and can alter the specificitybetween LCOs when using regions from a high affinity and specificity LCOLysM receptor such as a legume NFR1 receptor.

Certain aspects of the present disclosure relate to a modified plantLysM receptor comprising a LysM2 domain modified to comprise ahydrophobic patch on the surface of the LysM2 domain. In someembodiments, the modified LysM2 domain binds a lipo-chitooligosaccharide(LCO). In some embodiments, the modified LysM2 domain binds the LCO withhigher affinity than the unmodified LysM2 domain. In some embodiments,the modified LysM2 domain binds the LCO with higher selectivity for theLCO than the unmodified LysM2 domain. In some embodiments, the higheraffinity or higher selectivity is due to the hydrophobic patchinteracting with the LCO. In some embodiments, the higher affinity orhigher selectivity is due to the hydrophobic patch interacting with thelipid of the LCO. In some embodiments, the LCO is produced bynitrogen-fixing bacteria or by mycorrhizal fungi. In some embodiments,the LCO is produced by nitrogen-fixing bacteria selected from the groupof Mesorhizobium loti, Mesorhizobium huakuii, Mesorhizobiummediterraneum, Mesorhizobium ciceri, Mesorhizobium spp., Rhizobiummongolense, Rhizobium tropici, Rhizobium etli phaseoli, Rhizobiumgiardinii, Rhizobium leguminosarum optionally R. leguminosarum trifolii,R. leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., and any combination thereof,or by mycorrhizal fungi selected from the group consisting ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, or any combination thereof.

In some embodiments of any of the above embodiments, the LysM receptoris selected from the group consisting of a LysM chitooligosaccharide(CO) receptor, a LysM LCO receptor, and a LysM peptidoglycan (PGN)receptor. In some embodiments, the hydrophobic patch is adjacent to achitin binding motif if the LysM receptor is the LysM CO receptor or theLysM LCO receptor. In some embodiments, the hydrophobic patch is within30 Å, 20 Å, 10 Å, 7.5 Å, 5 Å, 4 Å, 3 Å, 2 Å, 1.5 Å, or 1 Å of a chitinbinding motif if the LysM receptor is the LysM CO receptor or the LysMLCO receptor. In some embodiments, the hydrophobic patch is adjacent toa glycan binding motif if the LysM receptor is the LysM PGN receptor. Insome embodiments, the hydrophobic patch is within 30 Å, 20 Å, 10 Å, 7.5Å, 5 Å, 4 Å, 3 Å, 2 Å, 1.5 Å, or 1 Å of a glycan binding motif if theLysM receptor is the LysM PGN receptor. In some embodiments, the LysMreceptor is not an exopolysaccharide (EPS) receptor.

In some embodiments of any of the above embodiments, the LysM receptoris a polypeptide having at least 70% sequence identity, at least 75%sequence identity, at least 80% sequence identity, at least 85% sequenceidentity, at least 90% sequence identity, at least 95% sequenceidentity, at least 96% sequence identity, at least 97% sequenceidentity, at least 98% sequence identity, or at least 99% sequenceidentity to SEQ ID NO:34 (i.e., Lotus CERK6; BAI79273.1_LjCERK6). Insome embodiments of any of the above embodiments, the LysM receptor is apolypeptide having amino acid sequence SEQ ID NO:34 (i.e., Lotus CERK6;BAI79273.1_LjCERK6).

In some embodiments of any of the above embodiments, the hydrophobicpatch was generated by deleting at least one non-hydrophobic amino acidresidue, substituting at least one amino acid residue with a morehydrophobic amino acid, or combinations thereof. In some embodiments,the at least one amino acid was identified by an amino acid sequencealignment with a LysM2 domain from a LysM high affinity LCO receptorthat naturally has a hydrophobic patch that interacts with LCO. In someembodiments, the at least one amino acid corresponds to an amino acidthat is red highlighted in red in FIGS. 12A-12G and FIG. 13C. In someembodiments, the at least one amino acid corresponds to an amino acidthat is red highlighted in red in FIGS. 12A-12G and FIG. 13C orcorresponds to an amino acid that is immediately N-terminal orC-terminal to an amino acid that is red highlighted in red in FIGS.12A-12G and FIG. 13C. In some embodiments, the at least one amino acidcorresponds to an amino acid that is red highlighted in red in FIGS.12A-12G in a known LCO receptor. In some embodiments, the at least oneamino acid corresponds to an amino acid that is red highlighted in redin FIGS. 12A-12G in a known LCO receptor or corresponds to an amino acidthat is immediately N-terminal or C-terminal to an amino acid that isred highlighted in red in FIGS. 12A-12G in a known LCO receptor. In someembodiments, the at least one amino acid was identified by structuralmodelling to identify a region in LysM2 where the hydrophobic patch canbe engineered. In some embodiments, the structural modeling used theunmodified plant LysM amino acid sequence and a LysM domain threedimensional structure that has a known hydrophobic patch. In someembodiments, the LysM domain three dimensional structure is a MedicagoNFP ectodomain. In some embodiments, the known hydrophobic patch aminoacid residues of the LysM domain three dimensional structure are orcorrespond to L147, L151, L152, L154, T156, K157 and V158 of theMedicago NFP ectodomain. In some embodiments, the alpha carbon of atleast one amino acid was within 3 Å of an alpha carbon of a knownhydrophobic patch amino acid residue in the structural alignment. Insome embodiments, the structural modeling was performed usingSWISS-MODEL, PDB2PQR, APBS, PyMol, and APBS tools 2.1.

In some aspects, the present disclosure relates to a modified plant LysMreceptor comprising a first LysM1 domain modified to replace at leastpart of the first LysM1 domain with at least part of a second LysM1domain. In some embodiments, the first LysM1 domain is modified bysubstituting a first part of the first LysM1 domain with a third part ofa second LysM1 domain and/or by substituting a second part of the firstLysM1 domain with a fourth part of the second LysM1 domain. In someembodiments, the first LysM1 domain and the second LysM1 domain havedifferent affinities, selectivities, and/or specificities foroligosaccharides and the modification of the first LysM1 domain altersthe affinity, selectivity, and/or specificity to be more like the secondLysM1 domain. In some embodiments, the first part and the third partcorrespond to SEQ ID NO:30 [Lotus CERK6 region II 43-53] or NGSNLTYISEI,SEQ ID NO:28 [Lotus NFR1 region II 41-52] or PGVFILQNITTF; and whereinthe second part and the fourth part correspond to SEQ ID NO:31 [LotusCERK6 region IV 74-82] or ASKDSVQAG; SEQ ID NO:29 [Lotus NFR1 region IV73-81], or LNDINIQSF. In some embodiments, the first LysM1 domain isselected from the group of SEQ ID NO:32 [LysM1 domain Lotus NFR1;LjNFR1/26-95], SEQ ID NO:33 [LysM1 domain Medicago LYK3; MtLYK3/25-95],or NFR1 DLALASYYILPGVFILQNITTFMQSEIVSSNDAITSYNKDKILNDINIQSFQRLNIPFP; andthe second LysM1 domain is CERK6:ALAQASYYLLNGSNLTYISEIMQSSLLTKPEDIVSYNQDTIASKDSVQAGQRINVPFP. In someembodiments, the first part is selected from SEQ ID NO:30 [Lotus CERK6region II 43-53] or NGSNLTYISEI; the second part is selected from SEQ IDNO:28 [Lotus CERK6 region IV 74-82] or ASKDSVQAG; the third part isselected from SEQ ID NO:31 [Lotus NFR1 region II 41-52] or PGVFILQNITTF;and the fourth part is selected from SEQ ID NO:29 [Lotus NFR1 region IV73-81] or LNDINIQSF. In some embodiments, the entire first LysM1 domainwas replaced with the entire second LysM1 domain. In some embodiments,the modified LysM1 domain binds a lipo-chitooligosaccharide (LCO)produced by nitrogen-fixing bacteria or by mycorrhizal fungi. In someembodiments, the LCO is produced by nitrogen-fixing bacteria selectedfrom the group consisting of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., and any combinationthereof, or by mycorrhizal fungi selected from the group consisting ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, and any combination thereof. Insome embodiments, the modified LysM1 domain binds an LCO with higheraffinity than an unmodified LysM1 domain. In some embodiments, themodified LysM1 domain binds LCOs with higher selectivity than anunmodified LysM1 domain. In some embodiments, the modified LysM1 domainbinds LCOs with altered specificity as compared to an unmodified LysM1domain. In some embodiments, structural modelling was used to define theLysM1 domain and was used to identify the first part, the second part,the third part, and/or the fourth part for substitution. In someembodiments, the receptor of the above embodiments further contains aLysM2 domain modified to contain a hydrophobic patch as in any one ofthe previous embodiments relating to modifying the LysM2 domain.

In some aspects, the present disclosure relates to a genetically alteredplant or part thereof, comprising a nucleic acid sequence encoding themodified plant LysM receptor of any one of the preceding embodiments. Insome embodiments, the modified plant LysM receptor has higher affinity,higher selectivity, and/or altered specificity for LCOs than theunmodified plant LysM receptor and the expression of the modified plantLysM receptor allows the plant or part thereof to recognize LCOs withhigh affinity, high selectivity, and/or altered specificity. In someembodiments, the LCOs are produced by nitrogen-fixing bacteria or bymycorrhizal fungi. In some embodiments, the LCOs are produced bynitrogen-fixing bacteria selected from the group consisting ofMesorhizobium loti, Mesorhizobium huakuii, Mesorhizobium mediterraneum,Mesorhizobium ciceri, Mesorhizobium spp., Rhizobium mongolense,Rhizobium tropici, Rhizobium etli phaseoli, Rhizobium giardinii,Rhizobium leguminosarum optionally R. leguminosarum trifolii, R.leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., and any combination thereof,or by or by mycorrhizal fungi selected from the group consisting ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, and any combination thereof. Insome embodiments, the modified polypeptide is localized to a plant cellplasma membrane. In some embodiments, the plant cell is a root cell. Insome embodiments, the root cell is a root epidermal cell. In someembodiments, the modified polypeptide is expressed in a developing plantroot system. In some embodiments, the nucleic acid sequence is operablylinked to a promoter. In some embodiments, the promoter is a rootspecific promoter. In some embodiments, the promoter is selected fromthe group of a NFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5promoter (SEQ ID NO:24) and the Lotus NFR1 promoters (SEQ ID NO:25) themaize allothioneine promoter, the chitinase promoter, the maize ZRP2promoter, the tomato LeExt1 promoter, the glutamine synthetase soybeanroot promoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter, a derivative of the CaMV35Spromoter, the maize ubiquitin promoter, the trefoil promoter, a veinmosaic cassava virus promoter, or the Arabidopsis UBQ10 promoter. Insome embodiments, the plant is selected from the group of corn, rice,barley, wheat, Trema spp., apple, pear, plum, apricot, peach, almond,walnut, strawberry, raspberry, blackberry, red currant, black currant,melon, cucumber, pumpkin, squash, grape, bean, pea, chickpea, cowpea,pigeon pea, lentil, Bambara groundnut, lupin, pulses, Medicago spp.,Lotus spp., forage legumes, indigo, legume trees, or hemp. In someembodiments, the plant part is a leaf, a stem, a root, a root primordia,a flower, a seed, a fruit, a kernel, a grain, a cell, or a portionthereof.

In some aspects, the present disclosure relates to a genetically alteredplant or part thereof, comprising a first nucleic acid sequence encodinga modified plant LysM receptor where the LysM1 domain has been modifiedas in any of the preceding embodiments relating to modification to theLysM1 domain and a second nucleic acid sequence encoding a modifiedplant LysM receptor where the LysM2 domain has been modified to includea hydrophobic patch as in any of the preceding embodiments relating tomodifications to the LysM2 domain. In some embodiments, the modifiedplant LysM receptor has higher affinity, higher selectivity, and/oraltered specificity for LCOs than the unmodified plant LysM receptor andthe expression of the modified plant LysM receptor allows the plant orpart thereof to recognize LCOs with high affinity, high selectivity,and/or altered specificity. In some embodiments, the LCOs are producedby nitrogen-fixing bacteria or by mycorrhizal fungi. In someembodiments, the LCOs are produced by nitrogen-fixing bacteria selectedfrom the group consisting of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., and any combinationthereof, or by or by mycorrhizal fungi selected from the groupconsisting of Acaulosporaceae spp., Diversisporaceae spp., Gigasporaceaespp., Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagusspp., Sclerocystis spp., Septoglomus spp., Claroideoglomus spp.,Ambispora spp., Archaeospora spp., Geosiphon pyriformis, Paraglomusspp., other species in the division Glomeromycota, and any combinationthereof. In some embodiments, the modified polypeptide is localized to aplant cell plasma membrane. In some embodiments, the plant cell is aroot cell. In some embodiments, the root cell is a root epidermal cell.In some embodiments, the modified polypeptide is expressed in adeveloping plant root system. In some embodiments, the first nucleicacid or second nucleic acid sequence is operably linked to a promoter.In some embodiments, the promoter is a root specific promoter. In someembodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO: 24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter, a derivative of the CaMV35Spromoter, the maize ubiquitin promoter, the trefoil promoter, a veinmosaic cassava virus promoter, or the Arabidopsis UBQ10 promoter. Insome embodiments, the plant is selected from the group of corn, rice,barley, wheat, Trema spp., apple, pear, plum, apricot, peach, almond,walnut, strawberry, raspberry, blackberry, red currant, black currant,melon, cucumber, pumpkin, squash, grape, bean, pea, chickpea, cowpea,pigeon pea, lentil, Bambara groundnut, lupin, pulses, Medicago spp.,Lotus spp., forage legumes, indigo, legume trees, or hemp. In someembodiments, the plant part is a leaf, a stem, a root, a root primordia,a flower, a seed, a fruit, a kernel, a grain, a cell, or a portionthereof. In some embodiments, the plant part is a fruit, a kernel, or agrain.

In some aspects, the present disclosure relates to a pollen grain or anovule of a genetically altered plant of any of the above embodimentsrelating to plants.

In some aspects, the present disclosure relates to a protoplast from agenetically altered plant of any of the above embodiments relating toplants.

In some aspects, the present disclosure relates to a tissue cultureproduced from protoplasts or cells from a genetically altered plant ofany of the above embodiments relating to plants, wherein the cells orprotoplasts are produced from a plant part selected from the groupconsisting of leaf, anther, pistil, stem, petiole, root, root primordia,root tip, fruit, seed, flower, cotyledon, hypocotyl, embryo, andmeristematic cell.

In some aspects, the present disclosure relates to a method of producingthe genetically altered plant of any one of the above embodimentsrelating to plants, comprising introducing a genetic alteration to theplant comprising the nucleic acid sequence. In some embodiments, thenucleic acid sequence, the first nucleic acid sequence, and/or thesecond nucleic acid sequence is operably linked to a promoter. In someembodiments, the promoter is a root specific promoter. In someembodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO: 24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter (KAY et al. Science, 236, 4805,1987), a derivative of the CaMV35S promoter, the maize ubiquitinpromoter, the trefoil promoter, a vein mosaic cassava virus promoter, orthe Arabidopsis UBQ10 promoter. In some embodiments, the nucleic acidsequence, the first nucleic acid sequence, and/or the second nucleicacid sequence is inserted into the genome of the plant so that thenucleic acid sequence, the first nucleic acid sequence, and/or thesecond nucleic acid sequence is operably linked to an endogenouspromoter. In some embodiments, the endogenous promoter is a rootspecific promoter.

Additional aspects of the present disclosure relate to a modified plantLysM receptor including a LysM2 domain modified to include a hydrophobicpatch on the surface of the LysM2 domain. In some embodiments, themodified LysM2 domain binds a lipo-chitooligosaccharide (LCO). In someembodiments, the modified LysM2 domain binds the LCO with higheraffinity than the unmodified LysM2 domain. In some embodiments, themodified LysM2 domain binds the LCO with higher selectivity for the LCOthan the unmodified LysM2 domain. In some embodiments, the higheraffinity or higher selectivity is due to the hydrophobic patchinteracting with the LCO. In some embodiments, the higher affinity orhigher selectivity is due to the hydrophobic patch interacting with thelipid of the LCO. In some embodiments, the LCO is produced bynitrogen-fixing bacteria or by mycorrhizal fungi. In some embodiments,the LCO is produced by nitrogen-fixing bacteria selected from the groupof Mesorhizobium loti, Mesorhizobium huakuii, Mesorhizobiummediterraneum, Mesorhizobium ciceri, Mesorhizobium spp., Rhizobiummongolense, Rhizobium tropici, Rhizobium etli phaseoli, Rhizobiumgiardinii, Rhizobium leguminosarum optionally R. leguminosarum trifolii,R. leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., or any combination thereof,or by mycorrhizal fungi selected from the group of Acaulosporaceae spp.,Diversisporaceae spp., Gigasporaceae spp., Pacisporaceae spp.,Funneliformis spp., Glomus spp., Rhizophagus spp., Sclerocystis spp.,Septoglomus spp., Claroideoglomus spp., Ambispora spp., Archaeosporaspp., Geosiphon pyriformis, Paraglomus spp., other species in thedivision Glomeromycota, or any combination thereof.

In some embodiments of any of the above embodiments, the LysM receptoris selected from the group of a LysM chitooligosaccharide (CO) receptor,a LysM LCO receptor, or a LysM peptidoglycan (PGN) receptor. In someembodiments, the hydrophobic patch is adjacent to a chitin binding motifif the LysM receptor is the LysM CO receptor or the LysM LCO receptor.In some embodiments, the hydrophobic patch is within 30 Å, 20 Å, 10 Å,7.5 Å, 5 Å, 4 Å, 3 Å, 2 Å, 1.5 Å, or 1 Å of a chitin binding motif ifthe LysM receptor is the LysM CO receptor or the LysM LCO receptor. Insome embodiments, the hydrophobic patch is adjacent to a glycan bindingmotif if the LysM receptor is the LysM PGN receptor. In someembodiments, the hydrophobic patch is within 30 Å, 20 Å, 10 Å, 7.5 Å, 5Å, 4 Å, 3 Å, 2 Å, 1.5 Å, or 1 Å of a glycan binding motif if the LysMreceptor is the LysM PGN receptor. In some embodiments, the LysMreceptor is not an exopolysaccharide (EPS) receptor.

In some embodiments of any of the above embodiments, the LysM receptoris a polypeptide having at least 70% sequence identity, at least 75%sequence identity, at least 80% sequence identity, at least 85% sequenceidentity, at least 90% sequence identity, at least 95% sequenceidentity, at least 96% sequence identity, at least 97% sequenceidentity, at least 98% sequence identity, or at least 99% sequenceidentity to SEQ ID NO:34 (i.e., Lotus CERK6; BAI79273.1_LjCERK6). Insome embodiments of any of the above embodiments, the LysM receptor is apolypeptide having amino acid sequence SEQ ID NO:34 (i.e., Lotus CERK6;BAI79273.1_LjCERK6).

In some embodiments of any of the above embodiments, the hydrophobicpatch was generated by deleting at least one non-hydrophobic amino acidresidue, substituting at least one amino acid residue with a morehydrophobic amino acid, or combinations thereof. In some embodiments ofany of the above embodiments, the hydrophobic patch was generated bymodifying an existing hydrophobic patch in the unmodified LysM receptor.In some embodiments, the unmodified LysM receptor was modified bydeleting at least one non-hydrophobic amino acid residue, substitutingat least one amino acid residue with a more hydrophobic amino acid,substituting at least one hydrophobic amino acid residue with anotherhydrophobic amino acid residue, or combinations thereof. In someembodiments, the at least one amino acid was identified by an amino acidsequence alignment with a LysM2 domain from a LysM high affinity LCOreceptor that naturally has a hydrophobic patch that interacts with LCO.In some embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G and FIG. 13C. In someembodiments, the at least one amino acid corresponds to an amino acidthat is in bold underline in FIGS. 12A-12G and FIG. 13C or correspondsto an amino acid that is immediately N-terminal or C-terminal to anamino acid that is in bold underline in FIGS. 12A-12G and FIG. 13C. Insome embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G in a known LCO receptor.In some embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G in a known LCO receptoror corresponds to an amino acid that is immediately N-terminal orC-terminal to an amino acid that is in bold underline in FIGS. 12A-12Gin a known LCO receptor. In some embodiments, the at least one aminoacid was identified by structural modelling to identify a region inLysM2 where the hydrophobic patch can be engineered. In someembodiments, the structural modeling used the unmodified plant LysMamino acid sequence and a LysM domain three dimensional structure thathas a known hydrophobic patch. In some embodiments, the LysM domainthree dimensional structure is a Medicago NFP ectodomain. In someembodiments, the known hydrophobic patch amino acid residues of the LysMdomain three dimensional structure are or correspond to L147, L151,L152, L154, T156, K157 and V158 of the Medicago NFP ectodomain. In someembodiments, the LysM domain three dimensional structure is a LotusLYS11 ectodomain. In some embodiments, the unmodified LysM receptor isthe Lotus LYS11 receptor and the existing hydrophobic patch amino acidresidues of the LysM domain that are modified are or correspond to K100,E101, G102, E103, S104, Y105, Y106, N128, and Y129 of the Lotus LYS11ectodomain. In some embodiments, the alpha carbon of at least one aminoacid was within 3 Å of an alpha carbon of a known hydrophobic patchamino acid residue in the structural alignment. In some embodiments, thestructural modeling was performed using SWISS-MODEL, PDB2PQR, APBS,PyMol, and APBS tools 2.1. In some embodiments of any of the aboveembodiments, either or both (i) 80% or fewer, 70% or fewer, 60% orfewer, 50% or fewer, 40% or fewer, 30% or fewer, or 20% or fewer ofamino acid residues in the LysM2 domain of the unmodified LysM receptorwere substituted or deleted to generate the modified plant LysMreceptor, and (ii) the entire LysM2 domain in the unmodified plant LysMreceptor was not substituted with another entire LysM2 domain togenerate the modified plant LysM receptor. In some embodiments of any ofthe above embodiments, the unmodified plant LysM receptor was selectedusing the method of any one of the aspects of the present disclosurerelating to such selection including any and all embodiments thereof.

In some aspects, the present disclosure relates to a modified plant LysMreceptor including a first LysM1 domain modified to replace at leastpart of the first LysM1 domain with at least part of a second LysM1domain. In some embodiments, the first LysM1 domain is modified bysubstituting a first part of the first LysM1 domain with a third part ofa second LysM1 domain and/or by substituting a second part of the firstLysM1 domain with a fourth part of the second LysM1 domain. In someembodiments, the first LysM1 domain and the second LysM1 domain havedifferent affinities, selectivities, and/or specificities foroligosaccharides and the modification of the first LysM1 domain altersthe affinity, selectivity, and/or specificity to be more like the secondLysM1 domain. In some embodiments, the first part and the third partcorrespond to SEQ ID NO:30 [Lotus CERK6 region II 43-53] or NGSNLTYISEI,SEQ ID NO:28 [Lotus NFR1 region II 41-52] or PGVFILQNITTF; and whereinthe second part and the fourth part correspond to SEQ ID NO:31 [LotusCERK6 region IV 74-82] or ASKDSVQAG; SEQ ID NO:29 [Lotus NFR1 region IV73-81], or LNDINIQSF. In some embodiments, the first LysM1 domain isselected from the group of SEQ ID NO:32 [LysM1 domain Lotus NFR1;LjNFR1/26-95], SEQ ID NO:33 [LysM1 domain Medicago LYK3; MtLYK3/25-95],or NFR1 DLALASYYILPGVFILQNITTFMQSEIVSSNDAITSYNKDKILNDINIQSFQRLNIPFP; andthe second LysM1 domain is CERK6:ALAQASYYLLNGSNLTYISEIMQSSLLTKPEDIVSYNQDTIASKDSVQAGQRINVPFP. In someembodiments, the first part is selected from SEQ ID NO:30 [Lotus CERK6region II 43-53] or NGSNLTYISEI; the second part is selected from SEQ IDNO:28 [Lotus CERK6 region IV 74-82] or ASKDSVQAG; the third part isselected from SEQ ID NO:31 [Lotus NFR1 region II 41-52] or PGVFILQNITTF;and the fourth part is selected from SEQ ID NO:29 [Lotus NFR1 region IV73-81] or LNDINIQSF. In some embodiments of any of the above embodimentsincluding the first LysM1 domain being modified to replace at least partof the first LysM1 domain with at least part of a second LysM1 domain,the first LysM1 domain is further modified by substituting a fifth partof the first LysM1 domain with a sixth part of a second LysM1 domain. Insome embodiments, the first LysM1 domain is SEQ ID NO:115 [LysM1 domainLotus NFR1; LjNFR1/32-89] or SEQ ID NO:106 [LysM1 domain Lotus NFR1;LjNFR1/31-89] and the second LysM1 domain is SEQ ID NO:114 [LysM1 domainMedicago LYK3; MtLYK3/31-89] or SEQ ID NO:105 [LysM1 domain MedicagoLYK3; MtLYK3/30-89]. In some embodiments, wherein the fifth part is SEQID NO:53 [Lotus NFR1 region III 59-62; LjNFR1/56-92], and wherein thesixth part is SEQ ID NO:46 [Medicago LYK3 region III 57-62;MtLYK3/57-62]. In some embodiments, the first LysM1 domain is modifiedby substituting a seventh part of the first LysM1 domain, wherein theseventh part spans the first part of the first LysM1 domain, the secondpart of the first LysM1 domain, and the fifth part of the first LysM1domain, with an eighth part of the second LysM1 domain, wherein theeighth part spans the third part of the second LysM1 domain, the fourthpart of the second LysM1 domain, and the sixth part of the second LysM1domain. In some embodiments, the seventh part of the first LysM1 domainis SEQ ID NO:51 [Lotus NFR1 regions II-IV 41-82; LjNFR1/41-82], and theeighth part of the second LysM1 domain is SEQ ID NO:113 [Medicago LYK3regions II-IV 40-82; MtLYK3/40-82] or SEQ ID NO:104 [Medicago LYK3regions II-IV 41-82; MtLYK3/41-82]. In some embodiments, the first LysM1domain is SEQ ID NO:33 [LysM1 domain Medicago LYK3; MtLYK3/31-89] andthe second LysM1 domain is SEQ ID NO:32 [LysM1 domain Lotus NFR1;LjNFR1/32-89]. In some embodiments, the fifth part is SEQ ID NO:46[Medicago LYK3 region III 57-62; MtLYK3/57-62], and the sixth part isSEQ ID NO:53 [Lotus NFR1 region III 59-62; LjNFR1/59-62]. In someembodiments of any of the above embodiments including the first LysM1domain being SEQ ID NO:33 and the second LysM1 domain being SEQ IDNO:32, the first LysM1 domain is modified by substituting a seventh partof the first LysM1 domain, wherein the seventh part spans the first partof the first LysM1 domain, the second part of the first LysM1 domain,and the fifth part of the first LysM1 domain, with an eighth part of thesecond LysM1 domain, wherein the eighth part spans the third part of thesecond LysM1 domain, the fourth part of the second LysM1 domain, and thesixth part of the second LysM1 domain. In some embodiments, the seventhpart of the first LysM1 domain is SEQ ID NO:51 [Lotus NFR1 regions II-IV41-82; LjNFR1/41-82], and the eighth part of the second LysM1 domain isSEQ ID NO:113 [Medicago LYK3 regions II-IV 40-82; MtLYK3/40-82] or SEQID NO:104 [Medicago LYK3 regions II-IV 41-82; MtLYK3/41-82].

In some embodiments of any of the above embodiments including the firstLysM1 domain being modified to replace at least part of the first LysM1domain with at least part of a second LysM1 domain, the entire firstLysM1 domain was replaced with the entire second LysM1 domain. In someembodiments of any of the above embodiments including the first LysM1domain being modified to replace at least part of the first LysM1 domainwith at least part of a second LysM1 domain, either or both (i) 80% orfewer, 70% or fewer, 60% or fewer, 50% or fewer, 40% or fewer, 30% orfewer, or 20% or fewer of amino acid residues in the first LysM1 domainwere substituted or deleted with the corresponding amino acid residuesof the second LysM1 domain, and (ii) the entire LysM1 domain in theunmodified plant LysM receptor was not substituted with another entireLysM2 domain to generate the modified plant LysM receptor. In someembodiments, the modified LysM1 domain binds a lipo-chitooligosaccharide(LCO) produced by nitrogen-fixing bacteria or by mycorrhizal fungi. Insome embodiments, the LCO is produced by nitrogen-fixing bacteriaselected from the group of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., or any combinationthereof, or by mycorrhizal fungi selected from the group ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, or any combination thereof. Insome embodiments, the modified LysM1 domain binds an LCO with higheraffinity than an unmodified LysM1 domain. In some embodiments, themodified LysM1 domain binds LCOs with higher selectivity than anunmodified LysM1 domain. In some embodiments, the modified LysM1 domainbinds LCOs with altered specificity as compared to an unmodified LysM1domain. In some embodiments, structural modelling was used to define theLysM1 domain and was used to identify the first part, the second part,the third part, and/or the fourth part for substitution. In someembodiments, the unmodified plant LysM receptor was selected using themethod of any one of the aspects of the present disclosure relating tosuch selection including any and all embodiments thereof and the secondLysM2 domain is from the donor plant LysM receptor. In some embodiments,the receptor of the above embodiments further contains a LysM2 domainmodified to contain a hydrophobic patch as in any one of the previousembodiments relating to modifying the LysM2 domain.

In some aspects, the present disclosure relates to a genetically alteredplant or part thereof, including a nucleic acid sequence encoding themodified plant LysM receptor of any one of the preceding embodiments. Insome embodiments, the modified plant LysM receptor has higher affinity,higher selectivity, and/or altered specificity for LCOs than theunmodified plant LysM receptor and the expression of the modified plantLysM receptor allows the plant or part thereof to recognize LCOs withhigh affinity, high selectivity, and/or altered specificity. In someembodiments, the LCOs are produced by nitrogen-fixing bacteria or bymycorrhizal fungi. In some embodiments, the LCOs are produced bynitrogen-fixing bacteria selected from the group of Mesorhizobium loti,Mesorhizobium huakuii, Mesorhizobium mediterraneum, Mesorhizobiumciceri, Mesorhizobium spp., Rhizobium mongolense, Rhizobium tropici,Rhizobium etli phaseoli, Rhizobium giardinii, Rhizobium leguminosarumoptionally R. leguminosarum trifolii, R. leguminosarum viciae, and R.leguminosarum phaseoli, Burkholderiales optionally symbionts of Mimosa,Sinorhizobium meliloti, Sinorhizobium medicae, Sinorhizobium fredii,Sinorhizobium NGR234, Azorhizobium caulinodans, Bradyrhizobiumjaponicum, Bradyrhizobium elkanii, Bradyrhizobium liaonginense, Frankiaspp., or any combination thereof, or by or by mycorrhizal fungi selectedfrom the group of Acaulosporaceae spp., Diversisporaceae spp.,Gigasporaceae spp., Pacisporaceae spp., Funneliformis spp., Glomus spp.,Rhizophagus spp., Sclerocystis spp., Septoglomus spp., Claroideoglomusspp., Ambispora spp., Archaeospora spp., Geosiphon pyriformis,Paraglomus spp., other species in the division Glomeromycota, or anycombination thereof. In some embodiments, the modified polypeptide islocalized to a plant cell plasma membrane. In some embodiments, theplant cell is a root cell. In some embodiments, the root cell is a rootepidermal cell. In some embodiments, the modified polypeptide isexpressed in a developing plant root system. In some embodiments, thenucleic acid sequence is operably linked to a promoter. In someembodiments, the promoter is a root specific promoter. In someembodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO:24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter, a derivative of the CaMV35Spromoter, the maize ubiquitin promoter, the trefoil promoter, a veinmosaic cassava virus promoter, or the Arabidopsis UBQ10 promoter. Insome embodiments, the plant is selected from the group of corn, rice,barley, wheat, Trema spp., apple, pear, plum, apricot, peach, almond,walnut, strawberry, raspberry, blackberry, red currant, black currant,melon, cucumber, pumpkin, squash, grape, bean, soybean, pea, chickpea,cowpea, pigeon pea, lentil, Bambara groundnut, lupin, pulses, Medicagospp., Lotus spp., forage legumes, indigo, legume trees, or hemp. In someembodiments, the plant part is a leaf, a stem, a root, a root primordia,a flower, a seed, a fruit, a kernel, a grain, a cell, or a portionthereof.

In some aspects, the present disclosure relates to a genetically alteredplant or part thereof, including a first nucleic acid sequence encodinga modified plant LysM receptor where the LysM1 domain has been modifiedas in any of the preceding embodiments relating to modification to theLysM1 domain and a second nucleic acid sequence encoding a modifiedplant LysM receptor where the LysM2 domain has been modified to includea hydrophobic patch as in any of the preceding embodiments relating tomodifications to the LysM2 domain. In some embodiments, the modifiedplant LysM receptor has higher affinity, higher selectivity, and/oraltered specificity for LCOs than the unmodified plant LysM receptor andthe expression of the modified plant LysM receptor allows the plant orpart thereof to recognize LCOs with high affinity, high selectivity,and/or altered specificity. In some embodiments, the LCOs are producedby nitrogen-fixing bacteria or by mycorrhizal fungi. In someembodiments, the LCOs are produced by nitrogen-fixing bacteria selectedfrom the group of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., or any combinationthereof, or by or by mycorrhizal fungi selected from the group ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, or any combination thereof. Insome embodiments, the modified polypeptide is localized to a plant cellplasma membrane. In some embodiments, the plant cell is a root cell. Insome embodiments, the root cell is a root epidermal cell. In someembodiments, the modified polypeptide is expressed in a developing plantroot system. In some embodiments, the first nucleic acid or secondnucleic acid sequence is operably linked to a promoter. In someembodiments, the promoter is a root specific promoter. In someembodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO: 24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter, a derivative of the CaMV35Spromoter, the maize ubiquitin promoter, the trefoil promoter, a veinmosaic cassava virus promoter, or the Arabidopsis UBQ10 promoter. Insome embodiments, the plant is selected from the group of corn, rice,barley, wheat, Trema spp., apple, pear, plum, apricot, peach, almond,walnut, strawberry, raspberry, blackberry, red currant, black currant,melon, cucumber, pumpkin, squash, grape, bean, soybean, pea, chickpea,cowpea, pigeon pea, lentil, Bambara groundnut, lupin, pulses, Medicagospp., Lotus spp., forage legumes, indigo, legume trees, or hemp. In someembodiments, the plant part is a leaf, a stem, a root, a root primordia,a flower, a seed, a fruit, a kernel, a grain, a cell, or a portionthereof. In some embodiments, the plant part is a fruit, a kernel, or agrain.

In some aspects, the present disclosure relates to a pollen grain or anovule of a genetically altered plant of any of the above embodimentsrelating to plants.

In some aspects, the present disclosure relates to a protoplast from agenetically altered plant of any of the above embodiments relating toplants.

In some aspects, the present disclosure relates to a tissue cultureproduced from protoplasts or cells from a genetically altered plant ofany of the above embodiments relating to plants, wherein the cells orprotoplasts are produced from a plant part selected from the group ofleaf, anther, pistil, stem, petiole, root, root primordia, root tip,fruit, seed, flower, cotyledon, hypocotyl, embryo, or meristematic cell.

In some aspects, the present disclosure relates to a method of producingthe genetically altered plant of any one of the above embodimentsrelating to plants, including introducing a genetic alteration to theplant having the nucleic acid sequence. In some embodiments, the nucleicacid sequence, the first nucleic acid sequence, and/or the secondnucleic acid sequence is operably linked to a promoter. In someembodiments, the promoter is a root specific promoter. In someembodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO: 24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter (KAY et al. Science, 236, 4805,1987), a derivative of the CaMV35S promoter, the maize ubiquitinpromoter, the trefoil promoter, a vein mosaic cassava virus promoter, orthe Arabidopsis UBQ10 promoter. In some embodiments, the nucleic acidsequence, the first nucleic acid sequence, and/or the second nucleicacid sequence is inserted into the genome of the plant so that thenucleic acid sequence, the first nucleic acid sequence, and/or thesecond nucleic acid sequence is operably linked to an endogenouspromoter. In some embodiments, the endogenous promoter is a rootspecific promoter.

In further aspects, the present disclosure relates to methods forselection of a target plant LysM receptor for modifying the target plantLysM receptor to have a desired receptor characteristic, wherein themethod includes the steps of: a) providing a structural model, amolecular model, a surface characteristics model, and/or anelectrostatic potential model of a donor plant LysM receptor having thedesired receptor characteristic and two or more potential target plantLysM receptors; b) comparing each of the two or more potential targetplant LysM receptors with the structural model, the molecular model, thesurface characteristics model, and/or the electrostatic potential modelof the donor plant LysM receptor, and/or comparing each of the two ormore potential target plant LysM receptors with the donor plant LysMreceptor using structural overlay; and c) selecting the potential targetplant LysM receptor with a suitable match for the donor plant LysMreceptor to be the target plant LysM receptor. In some embodiments, thecriteria for determining that the potential target plant LysM receptoris a suitable match for the donor plant LysM receptor in step (c) areselected from the group of goodness of fit to template structure;similarity; phylogenetic relation; surface potential; coverage totemplate structure; GMQE, QMEAN, and Local Quality estimates fromSWISS-Model; or any combination thereof. In some embodiments, thestructural model of a donor plant LysM receptor is a protein crystalstructure, a molecular model, a cryo-EM structure, and a NMR structure.In some embodiments, the donor plant LysM receptor model is of an entireectodomain and the two or more potential target plant LysM receptormodels are of entire ectodomains. In some embodiments, the donor plantLysM receptor model is of a LysM1 domain, a LysM2 domain, a LysM3domain, or any combination thereof, and the two or more potential targetplant LysM receptor models are of LysM1 domains, LysM2 domains, LysM3domains, or any combination thereof.

In some embodiments, the donor plant LysM receptor is Medicago NFP,Medicago LYK3, Lotus NFR1, Lotus NFR5, Lotus LYS11, or ArabidopsisCERK1. In some embodiments, the two or more target plant LysM receptorsare additionally compared to Lotus CERK6. In some embodiments, the twoor more potential target plant LysM receptor polypeptides are all fromthe same plant species or plant variety. In some embodiments, thedesired receptor characteristic is affinity, selectivity, and/orspecificity for an oligosaccharide or class of oligosaccharides. In someembodiments, the desired receptor characteristic is binding kinetics foran oligosaccharide or class of oligosaccharides, wherein the bindingkinetics include off-rate and on-rate. In some embodiments, the class ofoligosaccharides is selected from the group of LCOs, COs, beta-glucans,cyclic-beta-glucans, exopolysaccharides, or optionally LPS. In someembodiments, the class of oligosaccharides is LCOs or COs. In someembodiments, the class of oligosaccharides is LCOs, optionally producedby a produced by a nitrogen-fixing bacteria optionally selected from thegroup of Mesorhizobium loti, Mesorhizobium huakuii, Mesorhizobiummediterraneum, Mesorhizobium ciceri, Mesorhizobium spp., Rhizobiummongolense, Rhizobium tropici, Rhizobium etli phaseoli, Rhizobiumgiardinii, Rhizobium leguminosarum optionally R. leguminosarum trifolii,R. leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., or any combination thereof;or optionally produced by a mycorrhizal fungi optionally selected fromthe group of Acaulosporaceae spp., Diversisporaceae spp., Gigasporaceaespp., Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagusspp., Sclerocystis spp., Septoglomus spp., Claroideoglomus spp.,Ambispora spp., Archaeospora spp., Geosiphon pyriformis, Paraglomusspp., other species in the division Glomeromycota, pr any combinationthereof. In some embodiments, the LCOs are M. loti LCO, S. melilotiLCO-IV, or S. meliloti LCO-V.

In some embodiments, the method further includes step d) identifying oneor more amino acid residues for modification in the target LysM receptorby comparing amino acid residues of a first oligosaccharide bindingfeature in the donor plant LysM receptor with the corresponding aminoacid residues in the target plant LysM receptor, and optionallyidentifying one or more amino acid residues for modification in thetarget LysM receptor by comparing amino acid residues of a secondoligosaccharide binding feature in the donor plant LysM receptor withthe corresponding amino acid residues in the target plant LysM receptor.In some embodiments, the method further includes step e) generating amodified plant LysM receptor wherein the one or more amino acid residuesin the first oligosaccharide binding feature of the target plant LysMreceptor have been substituted with corresponding amino acid residuesfrom the donor LysM; generating a modified plant LysM receptor whereinthe one or more amino acid residues in the second oligosaccharidebinding feature of the target plant LysM receptor have been substitutedwith corresponding amino acid residues from the donor LysM; orgenerating a modified plant LysM receptor wherein the one or more aminoacid residues in the first oligosaccharide binding feature of the targetplant LysM receptor have been substituted with corresponding amino acidresidues from the donor LysM and the one or more amino acid residues inthe second oligosaccharide binding feature of the target plant LysMreceptor have been substituted with corresponding amino acid residuesfrom the donor LysM. In some embodiments, the first oligosaccharidebinding feature is a hydrophobic patch on the surface of the LysM2domain. In some embodiments, the second oligosaccharide binding featureis a part of the LysM1 domain of the donor plant LysM receptor.

In additional aspects, the present disclosure relates to a a modifiedplant LysM receptor produced using any one of the preceding methods,wherein the modified plant LysM receptor includes a LysM2 domainmodified to include a hydrophobic patch on the surface of the LysM2domain. In some embodiments, the modified LysM2 domain binds alipo-chitooligosaccharide (LCO). In some embodiments, the modified LysM2domain binds the LCO with higher affinity than the unmodified LysM2domain. In some embodiments, the modified LysM2 domain binds the LCOwith higher selectivity for the LCO than the unmodified LysM2 domain. Insome embodiments, the higher affinity or higher selectivity is due tothe hydrophobic patch interacting with the LCO. In some embodiments, thehigher affinity or higher selectivity is due to the hydrophobic patchinteracting with the lipid of the LCO. In some embodiments, the LCO isproduced by nitrogen-fixing bacteria or by mycorrhizal fungi. In someembodiments, the LCO is produced by nitrogen-fixing bacteria selectedfrom the group of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., and any combinationthereof, or by mycorrhizal fungi selected from the group ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, or any combination thereof.

In some embodiments of any of the above embodiments, the LysM receptoris selected from the group of a LysM chitooligosaccharide (CO) receptor,a LysM LCO receptor, or a LysM peptidoglycan (PGN) receptor. In someembodiments, the hydrophobic patch is adjacent to a chitin binding motifif the LysM receptor is the LysM CO receptor or the LysM LCO receptor.In some embodiments, the hydrophobic patch is within 30 Å, 20 Å, 10 Å,7.5 Å, 5 Å, 4 Å, 3 Å, 2 Å, 1.5 Å, or 1 Å of a chitin binding motif ifthe LysM receptor is the LysM CO receptor or the LysM LCO receptor. Insome embodiments, the hydrophobic patch is adjacent to a glycan bindingmotif if the LysM receptor is the LysM PGN receptor. In someembodiments, the hydrophobic patch is within 30 Å, 20 Å, 10 Å, 7.5 Å, 5Å, 4 Å, 3 Å, 2 Å, 1.5 Å, or 1 Å of a glycan binding motif if the LysMreceptor is the LysM PGN receptor. In some embodiments, the LysMreceptor is not an exopolysaccharide (EPS) receptor.

In some embodiments of any of the above embodiments, the LysM receptoris a polypeptide having at least 70% sequence identity, at least 75%sequence identity, at least 80% sequence identity, at least 85% sequenceidentity, at least 90% sequence identity, at least 95% sequenceidentity, at least 96% sequence identity, at least 97% sequenceidentity, at least 98% sequence identity, or at least 99% sequenceidentity to SEQ ID NO:34 (i.e., Lotus CERK6; BAI79273.1_LjCERK6). Insome embodiments of any of the above embodiments, the LysM receptor is apolypeptide having amino acid sequence SEQ ID NO:34 (i.e., Lotus CERK6;BAI79273.1_LjCERK6).

In some embodiments of any of the above embodiments, the hydrophobicpatch was generated by deleting at least one non-hydrophobic amino acidresidue, substituting at least one amino acid residue with a morehydrophobic amino acid, or combinations thereof. In some embodiments ofany of the above embodiments, the hydrophobic patch was generated bymodifying an existing hydrophobic patch in the unmodified LysM receptor.In some embodiments, the unmodified LysM receptor was modified bydeleting at least one non-hydrophobic amino acid residue, substitutingat least one amino acid residue with a more hydrophobic amino acid,substituting at least one hydrophobic amino acid residue with anotherhydrophobic amino acid residue, or combinations thereof. In someembodiments, the at least one amino acid was identified by an amino acidsequence alignment with a LysM2 domain from a LysM high affinity LCOreceptor that naturally has a hydrophobic patch that interacts with LCO.In some embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G and FIG. 13C. In someembodiments, the at least one amino acid corresponds to an amino acidthat is in bold underline in FIGS. 12A-12G and FIG. 13C or correspondsto an amino acid that is immediately N-terminal or C-terminal to anamino acid that is in bold underline in FIGS. 12A-12G and FIG. 13C. Insome embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G in a known LCO receptor.In some embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G in a known LCO receptoror corresponds to an amino acid that is immediately N-terminal orC-terminal to an amino acid that is in bold underline in FIGS. 12A-12Gin a known LCO receptor. In some embodiments, the at least one aminoacid was identified by structural modelling to identify a region inLysM2 where the hydrophobic patch can be engineered. In someembodiments, the structural modeling used the unmodified plant LysMamino acid sequence and a LysM domain three dimensional structure thathas a known hydrophobic patch. In some embodiments, the LysM domainthree dimensional structure is a Medicago NFP ectodomain. In someembodiments, the known hydrophobic patch amino acid residues of the LysMdomain three dimensional structure are or correspond to L147, L151,L152, L154, T156, K157 and V158 of the Medicago NFP ectodomain. In someembodiments, the LysM domain three dimensional structure is a LotusLYS11 ectodomain. In some embodiments, the unmodified LysM receptor isthe Lotus LYS11 receptor and the existing hydrophobic patch amino acidresidues of the LysM domain that are modified are or correspond to K100,E101, G102, E103, S104, Y105, Y106, N128, and Y129 of the Lotus LYS11ectodomain. In some embodiments, the alpha carbon of at least one aminoacid was within 3 Å of an alpha carbon of a known hydrophobic patchamino acid residue in the structural alignment. In some embodiments, thestructural modeling was performed using SWISS-MODEL, PDB2PQR, APBS,PyMol, and APBS tools 2.1. In some embodiments of any of the aboveembodiments, either or both (i) 80% or fewer, 70% or fewer, 60% orfewer, 50% or fewer, 40% or fewer, 30% or fewer, or 20% or fewer ofamino acid residues in the LysM2 domain of the unmodified LysM receptorwere substituted or deleted to generate the modified plant LysMreceptor, and (ii) the entire LysM2 domain in the unmodified plant LysMreceptor was not substituted with another entire LysM2 domain togenerate the modified plant LysM receptor. In some embodiments that maybe combined with any of the preceding embodiments, the presentdisclosure related to a genetically altered plant or part thereofincluding the modified plant LysM receptor of any of the aboveembodiments.

In further aspects, the present disclosure relates to a a modified plantLysM receptor produced using any one of the preceding methods, whereinthe modified plant LysM receptor includes a first LysM1 domain modifiedto replace at least part of the first LysM1 domain with at least part ofa second LysM1 domain. In some embodiments, the first LysM1 domain ismodified by substituting a first part of the first LysM1 domain with athird part of a second LysM1 domain and/or by substituting a second partof the first LysM1 domain with a fourth part of the second LysM1 domain.In some embodiments, the first LysM1 domain and the second LysM1 domainhave different affinities, selectivities, and/or specificities foroligosaccharides and the modification of the first LysM1 domain altersthe affinity, selectivity, and/or specificity to be more like the secondLysM1 domain. In some embodiments, the first part and the third partcorrespond to SEQ ID NO:30 [Lotus CERK6 region II 43-53] or NGSNLTYISEI,SEQ ID NO:28 [Lotus NFR1 region II 41-52] or PGVFILQNITTF; and whereinthe second part and the fourth part correspond to SEQ ID NO:31 [LotusCERK6 region IV 74-82] or ASKDSVQAG; SEQ ID NO:29 [Lotus NFR1 region IV73-81], or LNDINIQSF. In some embodiments, the first LysM1 domain isselected from the group of SEQ ID NO:32 [LysM1 domain Lotus NFR1;LjNFR1/26-95], SEQ ID NO:33 [LysM1 domain Medicago LYK3; MtLYK3/25-95],or NFR1 DLALASYYILPGVFILQNITTFMQSEIVSSNDAITSYNKDKILNDINIQSFQRLNIPFP; andthe second LysM1 domain is CERK6:ALAQASYYLLNGSNLTYISEIMQSSLLTKPEDIVSYNQDTIASKDSVQAGQRINVPFP. In someembodiments, the first part is selected from SEQ ID NO:30 [Lotus CERK6region II 43-53] or NGSNLTYISEI; the second part is selected from SEQ IDNO:28 [Lotus CERK6 region IV 74-82] or ASKDSVQAG; the third part isselected from SEQ ID NO:31 [Lotus NFR1 region II 41-52] or PGVFILQNITTF;and the fourth part is selected from SEQ ID NO:29 [Lotus NFR1 region IV73-81] or LNDINIQSF. In some embodiments of any of the above embodimentsincluding the first LysM1 domain being modified to replace at least partof the first LysM1 domain with at least part of a second LysM1 domain,the first LysM1 domain is further modified by substituting a fifth partof the first LysM1 domain with a sixth part of a second LysM1 domain. Insome embodiments, the first LysM1 domain is SEQ ID NO:115 [LysM1 domainLotus NFR1; LjNFR1/32-89] or SEQ ID NO:106 [LysM1 domain Lotus NFR1;LjNFR1/31-89] and the second LysM1 domain is SEQ ID NO:114 [LysM1 domainMedicago LYK3; MtLYK3/31-89] or SEQ ID NO:105 [LysM1 domain MedicagoLYK3; MtLYK3/30-89]. In some embodiments, wherein the fifth part is SEQID NO:53 [Lotus NFR1 region III 59-62; LjNFR1/59-62], and wherein thesixth part is SEQ ID NO:46 [Medicago LYK3 region III 57-62;MtLYK3/57-62]. In some embodiments, the first LysM1 domain is modifiedby substituting a seventh part of the first LysM1 domain, wherein theseventh part spans the first part of the first LysM1 domain, the secondpart of the first LysM1 domain, and the fifth part of the first LysM1domain, with an eighth part of the second LysM1 domain, wherein theeighth part spans the third part of the second LysM1 domain, the fourthpart of the second LysM1 domain, and the sixth part of the second LysM1domain. In some embodiments, the seventh part of the first LysM1 domainis SEQ ID NO:51 [Lotus NFR1 regions II-IV 41-82; LjNFR1/41-82], and theeighth part of the second LysM1 domain is SEQ ID NO:113 [Medicago LYK3regions II-IV 40-82; MtLYK3/40-82] or SEQ ID NO:104 [Medicago LYK3regions II-IV 41-82; MtLYK3/41-82]. In some embodiments, the first LysM1domain is SEQ ID NO:33 [LysM1 domain Medicago LYK3; MtLYK3/31-89] andthe second LysM1 domain is SEQ ID NO:32 [LysM1 domain Lotus NFR1;LjNFR1/32-89]. In some embodiments, the fifth part is SEQ ID NO:46[Medicago LYK3 region III 57-62; MtLYK3/57-62], and the sixth part isSEQ ID NO:53 [Lotus NFR1 region III 59-62; LjNFR1/59-62]. In someembodiments of any of the above embodiments including the first LysM1domain being SEQ ID NO:33 and the second LysM1 domain being SEQ IDNO:32, the first LysM1 domain is modified by substituting a seventh partof the first LysM1 domain, wherein the seventh part spans the first partof the first LysM1 domain, the second part of the first LysM1 domain,and the fifth part of the first LysM1 domain, with an eighth part of thesecond LysM1 domain, wherein the eighth part spans the third part of thesecond LysM1 domain, the fourth part of the second LysM1 domain, and thesixth part of the second LysM1 domain. In some embodiments, the seventhpart of the first LysM1 domain is SEQ ID NO:51 [Lotus NFR1 regions II-IV41-82; LjNFR1/41-82], and the eighth part of the second LysM1 domain isSEQ ID NO:113 [Medicago LYK3 regions II-IV 40-82; MtLYK3/40-82] or SEQID NO:104 [Medicago LYK3 regions II-IV 41-82; MtLYK3/41-82].

In some embodiments of any of the above embodiments including the firstLysM1 domain being modified to replace at least part of the first LysM1domain with at least part of a second LysM1 domain, the entire firstLysM1 domain was replaced with the entire second LysM1 domain. In someembodiments of any of the above embodiments including the first LysM1domain being modified to replace at least part of the first LysM1 domainwith at least part of a second LysM1 domain, either or both (i) 80% orfewer, 70% or fewer, 60% or fewer, 50% or fewer, 40% or fewer, 30% orfewer, or 20% or fewer of amino acid residues in the first LysM1 domainwere substituted or deleted with the corresponding amino acid residuesof the second LysM1 domain, and (ii) the entire LysM1 domain in theunmodified plant LysM receptor was not substituted with another entireLysM2 domain to generate the modified plant LysM receptor. In someembodiments, the modified LysM1 domain binds a lipo-chitooligosaccharide(LCO) produced by nitrogen-fixing bacteria or by mycorrhizal fungi. Insome embodiments, the LCO is produced by nitrogen-fixing bacteriaselected from the group of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., or any combinationthereof, or by mycorrhizal fungi selected from the group ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, or any combination thereof Insome embodiments, the modified LysM1 domain binds an LCO with higheraffinity than an unmodified LysM1 domain. In some embodiments, themodified LysM1 domain binds LCOs with higher selectivity than anunmodified LysM1 domain. In some embodiments, the modified LysM1 domainbinds LCOs with altered specificity as compared to an unmodified LysM1domain. In some embodiments, structural modelling was used to define theLysM1 domain and was used to identify the first part, the second part,the third part, and/or the fourth part for substitution. In someembodiments, the receptor of the above embodiments further contains aLysM2 domain modified to contain a hydrophobic patch as in any one ofthe previous embodiments relating to modifying the LysM2 domain. In someembodiments that may be combined with any of the preceding embodiments,the present disclosure related to a genetically altered plant or partthereof including the modified plant LysM receptor of any of the aboveembodiments.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the structure of the NFP receptor ectodomain (NFP-ECD) withthe three LysM domains labeled (LysM1, LysM2, and LysM3). Motifs withinthe LysM domains are also labeled: LysM1 motifs=α1, α2, β1, and β2;LysM2 motifs=α3, α4, β3, and β4; and LysM3 motifs=α5, α6, β5, and β6.Glycosylations (di-GlcNAc cores are shown (projecting from α1 at upper;additional cores visible at center adjacent to β2 and β1 as well as atbottom left behind α4), and disulfide bridges are indicated with arrowsand labeled with the residue numbers (C47-C166; C39-C104; andC102-C164).

FIGS. 2A-2B show the hydrophobic patch in the Medicago NFP LysM2 domain,and binding assay measurements using mutants of important residueswithin the hydrophobic patch. FIG. 2A shows molecular docking of CO4(designated as “Ligand”) onto Medicago NFP shaded with electrostaticsurface potential. The hydrophobic patch is circled by a dashed blackline, and the locations of important residues L147 and L154 are shownusing arrows. The position of the LCO fatty-acid is depicted with adashed grey line. FIG. 2B shows binding assay measurements comparing awild type (WT) NFP (“NFP WT”) with an NFP mutated at residues 147 and154 (“NFP L147D L154D”; bold). The results shown for NFP WT are fromseven replicates and the results shown for NFP L147D L154D are from fourreplicates.

FIG. 3 shows the general schematic of the construct used for mutantcomplementation experiments. Designations are as follows: T-DNA leftborder=LB, T-DNA right border=RB, nuclear localized triple yellowfluorescent protein=tYFPnls, buffer sequence=buffer, constitutiveubiquitin promoter=pUbi, Nfr1 promoter=pNfr1, Cerk6 promoter=pCerk6. Thearrows indicate the directions of gene transcription.

FIGS. 4A-4B show complementation assays of Medicago nfp mutants. FIG. 4Ashows complementation tested by inoculation with S. meliloti strain2011. FIG. 4B shows complementation tested by inoculation with S.medicae. Columns represent the mean nodule numbers, while circlesrepresent the individual counts. Empty circles=Medicago A17 wild type;filled circles=Medicago nfp mutant; EVC=empty vector control; andWT=wild type. Error bars show the SEM. Different letters indicatesignificant differences between the samples (ANOVA, Tukey, P<0.05).

FIGS. 5A-5B show results of functional studies measuring nodulation anddefense using domain swaps between the Lotus japonicus (Lj) LCO receptorNFR1 and the Lotus japonicus (Lj) CO receptor CERK6. FIG. 5A showscomplementation experiments of a Lotus nfr1-1 single mutant withdifferent domain-swapped protein constructs. Nodules were counted onhairy root transformed L. japonicus nfr1-1 mutant roots after theindicated days post inoculation (dpi) with M. loti R7A. FIG. 5B showscomplementation of a Lotus cerk6 single mutant with differentdomain-swapped protein constructs. Ratios of CO8 and flg22 elicited ROSpeak values are plotted normalized to the wild type sample (Gifu;transformed with the empty vector) set as 1. In both FIGS. 5A and 5B,black dots represent individual transformed plants, and error bars showthe SEM. Different letters indicate significant differences among thesamples (ANOVA, Tukey, P<0.01). The compositions of the recombinantreceptors are shown by shading of the respective parts, with blackindicating LjNFR1 derived sequences and grey indicating LjCERK6 derivedsequences. Shaded bars indicate multiple amino acid long region swaps(black bars are LjNFR1 derived sequences; grey bars are LjCERK6 derivedsequences), while white bars indicate single amino acid mutationsintroducing a bulky amino acid (Trp) to the LysM structure. LysM domainsare labelled at the left (in FIG. 5A) or at the right (in FIG. 5B) ofthe recombinant receptors as LysM1, LysM2, and LysM3, and transmembraneand intracellular domains are labelled at the left (in FIG. 5A) or atthe right (in FIG. 5B) of the recombinant receptors as TM+IC.

FIG. 6 shows a 3D structure of the Lotus CERK6 ectodomain with the threeLysM domains labeled (LysM1, LysM2, and LysM3). Region II and region IVin LysM1 are labeled and shaded in light grey.

FIG. 7 shows complementation of Lotus japonicus nfr1-1 mutants withLjNFR1/MtLYK3 chimeras depicted at the bottom of the graph.Complementation was assayed by counting nodules formed per plant, whichis shown at the top of FIG. 7. Black dots represent individual plants,columns indicate the mean values, and error bars show the SEM. Differentletters indicate significant difference among the samples (ANOVA, Tukey,P<0.01). The schematics of the individual chimeric receptors tested areshown at the bottom of FIG. 7, with white indicating LjNFR1 domains,grey indicating MtLYK3 domains, and black indicating LjCERK6 domains(control). LysM domains are labelled as LysM1, LysM2, and LysM3;transmembrane and juxtamembrane domains are labelled as TM and JM; andthe kinase domain is labelled as K

FIGS. 8A-8C show an alignment of selected LysM receptors fromArabidopsis thaliana (At; AT3G21630_CERK1 (SEQ ID NO:75), AT1G77630_LYP3(SEQ ID NO:80), AT2G17120_LYP1 (SEQ ID NO:82)), Zea mays (Zm;ZM9_NP_001146346.1 (SEQ ID NO:72)), Hordeum vulgare (Hv;HvLysMRLK4_AK369594.1 (SEQ ID NO:73)), Medicago truncatula (Mt or Medtr;Mt_LYK9XP_003601376 (SEQ ID NO:69), Mt_LYK3_XP_003616958 (SEQ ID NO:71),Mt_LYK10_XP_003613165 (SEQ ID NO:77), Medtr5g042440.1 (SEQ ID NO:79)),Oryza sativa (Os; XP_015611967_OsCERK1 (SEQ ID NO:74), OsCeBiP (SEQ IDNO:81)) and Lotus japonicus (Lj; BAI79273.1_CERK6 (SEQ ID NO:34),CAE02590.1_NFR1 (SEQ ID NO:70), BAI79284.1_EPR3 (SEQ ID NO:76),CAE02597.1_NFR5 (SEQ ID NO:78)). NFR1 and NFR5 are Nod factor receptors,EPR3 is an exopolysaccharide receptor, AtLYP1 and AtLYP3 arepeptidoglycan receptors, AtCERK1, OsCERK1, OsCeBIP, CERK6 arechitooligosaccharide receptors. C(x)XXXC and CxC motifs flanking thethree LysM domains are shown. LysM1 (black line), LysM2 (grey line) andLysM3 (grey line) are shown. FIG. 8A shows the first two portions of thealignment including all of the LysM1 domain and part of the LysM2domain. FIG. 8B shows the third and fourth portions of the alignmentincluding the rest of the LysM2 domain and all of the LysM3 domain. FIG.8C shows the fifth portion of the alignment.

FIGS. 9A-9B show an alignment of selected LysM receptors fromArabidopsis thaliana (At; AT3G21630_CERK1 (SEQ ID NO:75)), Zea mays (Zm;XP_020399958_ZM1 (SEQ ID NO:20), XP_008652982.1_ZM5 (SEQ ID NO:21),AQK73561.1_ZM7 (SEQ ID NO:84), NP_001147981.1_ZM3 (SEQ ID NO:85),NP_001147941.2_ZM6 (SEQ ID NO:86), AQK58792.1_ZM4 (SEQ ID NO:87),ZM9_NP_001146346.1 (SEQ ID NO:72)), Hordeum vulgare (Hv; HORVU4Hr1G066170_HvLysMRLK10 (SEQ ID NO:19), AK357612_HvLysMRLK2 (SEQ ID NO:17),AK370300_HvLysmRLK1 (SEQ ID NO:16), AK372128_HvLysMRLK3 (SEQ ID NO:18),HvLysMRLK4_AK369594.1 (SEQ ID NO:73)), Oryza sativa (Os;XP_015611967_OsCERK1 (SEQ ID NO:74)), Medicago truncatula (Mt;XP_003613904.2_MtNFP (SEQ ID NO:83), MtLYK3_XP_003616958 (SEQ ID NO:71),MtLYK9XP_003601376 (SEQ ID NO:69)), and Lotus japonicus (Lj;CAE02590.1_NFR1 (SEQ ID NO:70), CAE02597.1NFR5 (SEQ ID NO:78),BAI79273.1_CERK6 (SEQ ID NO:34). LjNFR1, LjNFR5, MtLYK3 and MtNFP arefunctional Nod factor receptors, AtCERK1, OsCERK1, LjCERK6 arefunctional chitin receptors. C(x)XXXC and CxC motifs flanking the threeLysM domains are shown. LysM1 (black line), LysM2 (grey line) and LysM3(grey line) are shown. The number of “X” residues in the C(x)XXXC motiflocated before LysM1 varies between receptors and therefore the locationof LysM1 (black line) changes accordingly in the alignments in thisfigure and in successive figures. FIG. 9A shows the first and secondportions of the alignment including all of the LysM1 domain and part ofthe LysM2 domain. FIG. 9B shows the third and fourth portions of thealignment including the rest of the LysM2 domain and all of the LysM3domain.

FIGS. 10A-10B show an alignment of selected LysM receptors from Zea mays(Zm; ONM41523.1_ZM8 (SEQ ID NO:88), XP_008657477.1_ZM2 (SEQ ID NO:89),Zm00001d043516 ZM10 (SEQ ID NO:91)), Hordeum vulgare (Hv;MLOC_5489.2_HvLysM-RLK9 (SEQ ID NO:90), MLOC_18610.1_HvLysM-RLK8 (SEQ IDNO:92), MLOC_57536.1_HvLysM-RLK6 (SEQ ID NO:93)), Medicago truncatula(Mt; Mt_LYK10_XP_003613165 (SEQ ID NO:77), Mt_LYK3_XP_003616958 (SEQ IDNO:71), XP_003613904.2_MtNFP (SEQ ID NO:83)), and Lotus japonicus (Lj;BAI79284.1_EPR3 (SEQ ID NO:76), CAE02590.1_NFR1 (SEQ ID NO:70),CAE02597.1_NFR5 (SEQ ID NO:78)). LjNFR1, LjNFR5, MtLYK3 and MtNFP arefunctional Nod factor receptors, LjEPR3 is functional EPS receptor.C(x)XXXC and CxC motifs flanking the three LysM domains are shown. LysM1(black line), LysM2 (grey line) and LysM3 (grey line) are shown. FIG.10A shows the first, second, and third portions of the alignmentincluding all of the LysM1 domain and all of the LysM2 domain. FIG. 10Bshows the fourth, fifth, and sixth portions of the alignment includingall of the LysM3 domain.

FIGS. 11A-11B show an alignment of selected LysM receptors fromArabidopsis thaliana (At; AT1G21880.2_LYP2 (SEQ ID NO:94),AT1G77630_LYP3 (SEQ ID NO:80), AT2G17120_LYP1 (SEQ ID NO:82)), Oryzasativa (Os; OsCeBiP (SEQ ID NO:81)), and Lotus japonicus (Lj; LjLYP1(SEQ ID NO:95), LjLYP2 (SEQ ID NO:96), LjLYP3 (SEQ ID NO:97),CAE02590.1_NFR1 (SEQ ID NO:70), CAE02597.1_NFR5 (SEQ ID NO:78)). LjNFR1,LjNFR5, are functional Nod factor receptors, AtLYP2 and AtLYP3, are PGNreceptors, OsCeBiP is a functional chitin receptor. C(x)XXXC and CxCmotifs flanking the three LysM domains are shown. LysM1 (black line),LysM2 (grey line) and LysM3 (grey line) are shown. FIG. 11A shows thefirst, second, third, and fourth portions of the alignment including allof the LysM1 domain, all of the LysM2 domain, and all of the LysM3domain. FIG. 11B shows the fifth, sixth, and seventh portions of thealignment.

FIGS. 12A-12E show annotated amino acid sequences of previously knownLCO receptors and newly identified LCO receptors. FIG. 12A shows theannotation key; the LysM1 domain is shown with a dashed underline, theLysM2 domain is shown with a solid underline, the hydrophobic patchresidues are shown in bold, and the LysM3 domain is shown with residuesitalicized. Medicago NFP (MtNFP/1-595; SEQ ID NO:1), Lotus NFR5 (a knownLCO receptor; LjNFR5/1-595; SEQ ID NO:2), Pea SYM10 (a known LCOreceptor; Pea_SYM10/1-594; SEQ ID NO:3), and Soybean NFR5α (a known LCOreceptor; GmNFR5α/1-598 max; SEQ ID NO:4) are shown. FIG. 12B showsChickpea NFR5 (a new LCO receptor; ChickpeaNFR5/1-557 (Cicer arietinum);SEQ ID NO:5), Bean NFR5 (a new LCO receptor; BeanNFR5/1-597 (Phaseolusvulgaris); SEQ ID NO:7), Peanut NFR5 (a new LCO receptor;PeanutNFR5/1-595 [Arachis hypogaea subsp. hypogaea]; SEQ ID NO:9), andLotus LYS11 (a new LCO receptor; LjLYS11/1-591; SEQ ID NO:11). FIG. 12Cshows Medicago LYR1 (a new LCO receptor; MtLYR1/1-590; SEQ ID NO:12),Parasponia NFP1 (a new LCO receptor; PanNFP1/1-613; SEQ ID NO:13),Parasponia NFP2 (a known LCO receptor; PanNFP2/1-582; SEQ ID NO:14), andBarley receptor HvLysM-RLK1 (a new LCO receptor; HvLysM-RLK1 (AK370300);SEQ ID NO:16). FIG. 12D shows Barley receptor HvLysM-RLK2 (a new LCOreceptor; HvLysM-RLK2 (AK357612); SEQ ID NO:17), Barley receptorHvLysM-RLK3 AK372128 (a new LCO receptor; HvLysM-RLK3 AK372128; SEQ IDNO:18), Barley receptor HvLysM-RLK10 (a new LCO receptor; HvLysM-RLK10(HORVU4Hr1G066170); SEQ ID NO:19), and Maize receptor ZM1 (a new LCOreceptor; ZM1 (XP_020399958); SEQ ID NO:20). FIG. 12E shows Maizereceptor ZM5 (a new LCO receptor; ZM5 (XP_008652982.1); SEQ ID NO:21),Apple NFP5 (a new LCO receptor; XP_008338966.1 PREDICTED:serine/threonine receptor-like kinase NFP [Malus domestica]; SEQ IDNO:22), and Strawberry NFR5 (a new LCO receptor; XP_004300586.2PREDICTED: protein LYK5-like [Fragaria vesca subsp. vesca]; SEQ IDNO:23).

FIGS. 13A-13C show structural modelling of the HvLysM-RLK2/37-247LysM1-3 domains and selection of residues for modification to introducea hydrophobic patch. FIG. 13A shows the PyMol visualization of theLysM1-3 domains of the HvLysM-RLK2/37-247 model with the LysM1 domainlabeled and in dark grey, the LysM2 labeled and in light grey, and theLysM3 labeled and in light grey. FIG. 13B shows the electrostaticsurface potential of the model with chitin modeled in the bindinggroove. FIG. 13C has the amino acid sequence of the HvLysM-RLK2/37-247LysM1-3 domains with the LysM1 domain with a dashed underline, the LysM2domain with a solid underline, and the LysM3 domain with no underline,and the residues that can be modified to create the hydrophobic patch inbold.

FIG. 14 shows an alignment of the Lotus japonicus (Lj) NFR1 and Lotusjaponicus (Lj) CERK6 LysM1 domains. Viewed top-down, LjNFR1(NFR1_Lj2g2904690; SEQ ID NO:99) is shown in the top row, LjCERK6 (CERK6AB503687; SEQ ID NO:100) is shown in the second row, sequenceconservation is shown in the third row. Region II and region IV in theLysM1 domain are denoted by light grey arrows and labelled as “II.” or

FIGS. 15A-15F show L. japonicus LjLYS11 ectodomain model and crystalstructure, modified LjLYS11 ectodomains, and testing of modified LjLYS11ectodomains. FIG. 15A shows a comparison of the LjLYS11 ectodomain model(LYS11—model; left) with the crystal structure of the LjLYS11 ectodomain(LYS11—crystal structure; right). FIG. 15B shows schematics of modifiedLjLYS11 ectodomains (LjLYS11—LjNFR5 chimeras) used for testing. The topschematic shows an ectodomain with entirely LjLYS11 domains (black), themiddle schematic shows an ectodomain where the LysM2 domain from LjLYS11was replaced with the LysM2 domain from LjNFR5 (grey), and the bottomschematic shows an ectodomain where key residues from LjLYS11 werereplaced with key residues from LjNFR5 (grey) (N-terminus=N′; LysM1=M1;LysM2=M2; LysM3=M3; 6×HIS tag used for purification=6×HIS;C-terminus=C′). FIG. 15C shows the results of binding assays with theectodomain with entirely LjLYS11 components (ectodomain schematic shownat top with LjLYS11 domains in black; results of binding assays shown atbottom). The Kd is shown in the title of each graph (CO5 (Kd=11.4 μM),M. loti LCO (Kd=38.6 μM), and S. meliloti LCO (weak binding)). FIG. 15Dshows the results of binding assays with the ectodomain where LysM2 fromLjLYS11 was replaced with LysM2 from LjNFR5 (ectodomain schematic shownat top with LjLYS11 domains in black and LjNFR5 domains in grey; resultsof binding assays shown at bottom). FIG. 15E shows the results ofbinding assays with the ectodomain where key residues from LjLYS11 werereplaced with key residues from LjNFR5 (ectodomain schematic shown attop with LjLYS11 domains in black and LjNFR5 residues in grey; resultsof binding assays shown at bottom). For FIGS. 15C-15E, binding in nm isshown on the y-axes, time in seconds (s) is shown on the x-axes, and thetested molecules are shown in the titles of the graphs (C05, M. lotiLCO, and S. meliloti LCO). FIG. 15F shows complementation of L.japonicus nfr5 (Ljnfr5) mutants with LjNFR5/LjLYS11 chimeras depicted atthe bottom of the graph. Complementation was assayed by counting nodulesformed per plant, which is shown at the top of FIG. 15F. Black dotsrepresent individual plants, columns indicate the mean values, and errorbars show the SEM. Different letters indicate significant differenceamong the samples (ANOVA, Tukey, P<0.01). The schematics of theindividual chimeric ectodomains tested are shown at the bottom of FIG.15F, with light grey indicating LjNFR5 domains, grey indicating LjLYS11domains, and empty vector denoted by a label (LysM1, LysM2 and LysM3 areshown as boxes; transmembrane domain is shown as a wavy shape; kinasedomain is shown as an oval shape). Below the receptor schematics, thenumber of plants (Plant), the number of plants without nodules (neg),the number of plants with nodules (pos), and the frequency (freq) ofplants forming nodules when transformed with the depicted vector isprovided.

FIGS. 16A-16H show homology modelling of the barley RLK10 receptor(HvRLK10) ectodomain and of the barley RLK4 receptor (HvRLK4) ectodomainas well as results of binding experiments using the HvRLK10 ectodomainand the HvRLK4 ectodomain. FIG. 16A shows a schematic of the purifiedHvRLK10 ectodomain at the top (N-terminus=N′; LysM1=M1; LysM2=M2;LysM3=M3; 6×HIS tag used for purification=6×HIS; C-terminus=C′) and theresults of binding assays of HvRLK10 ectodomain with CO5 at the bottom.FIG. 16B shows homology modelling of the Barley receptor RLK10 (HvRLK10)ectodomain with surface representation shaded according to itselectrostatic potential. The hydrophobic patch is circled by a dashedblack line, and the ligand is shown at the top of the hydrophobic patch.FIG. 16C shows the results of binding assays of HvRLK10 ectodomain withM. loti LCO. FIG. 16D shows the results of binding assays of HvRLK10ectodomain with S. meliloti LCO. FIG. 16E shows a schematic of thepurified HvRLK4 ectodomain at the top (N-terminus=N′; LysM1=M1;LysM2=M2; LysM3=M3; 6×HIS tag used for purification=6×HIS;C-terminus=C′) and the results of binding assays of HvRLK4 ectodomainwith CO5 at the bottom. FIG. 16F shows a 3D structure of the HvRLK4ectodomain with a ligand shown at the top. FIG. 16G shows the results ofbinding assays of HvRLK4 ectodomain with M. loti LCO. FIG. 16H shows theresults of binding assays of HvRLK4 ectodomain with S. meliloti LCO. ForFIGS. 16A, 16C-16D, 16E, and 16G-16H, binding in nm is shown on they-axes, time in seconds (s) is shown on the x-axes, and the testedmolecules are shown in the titles of the graphs (CO5, M. loti LCO, andS. meliloti LCO).

FIGS. 17A-17B show complementation of Medicago truncatula lyk3 mutantswith LjNFR1/MtLYK3 chimeras as well as the MtLYK3 ectodomain structureand schematics of LjNFR1/MtLYK3 chimeras. FIG. 17A shows complementationof Medicago lyk3 mutants (Mtlyk3) with LjNFR1/MtLYK3 chimeras depictedat the bottom of the graph. The controls Medicago wild type (WT)transformed with empty vector and Mtlyk3 transformed with empty vectorwere included. Complementation was assayed by counting nodules formedper plant, which is shown at the top of FIG. 17A. Black dots representindividual plants, columns indicate the mean values, and error bars showthe SEM. Different letters indicate significant difference among thesamples (ANOVA, Tukey, P<0.01). The schematics of the individualchimeric receptors tested are shown at the bottom of FIG. 17A, with greyindicating MtLYK3 domains, black indicating LjNFR1 domains, and emptyvector denoted by a label. Below the receptor schematics, the frequency(freq) of plants forming nodules when transformed with the depictedvector is provided. LysM domains are labelled as LysM1, LysM2, andLysM3; transmembrane domain is labelled as TM; and the kinase domain islabelled as kinase. FIG. 17B shows the MtLYK3 ectodomain structure, withRegion II (dark grey helix), Region III (light grey helix), and RegionIV (grey linker) labeled (top); and schematic representation of theengineered LjNFR1 ectodomain (black) with indicated regions from theMtLYK3 ectodomain (grey; regions from the MtLYK3 ectodomain=regionsII-IV (“2 to 4”), regions II, III, and IV (“2, 3, 4”), and regions IIand IV (“2, 4”)) (bottom).

FIGS. 18A-18F show BLI binding curves for A. thaliana CERK1 (AtCERK1)binding to chitopentaose (CO5) and chitooctaose (CO8), models of CO andLCO perception, and structural alignment of the ectodomains of MedicagoNFP, Arabidopsis CERK1 and Lotus CERK6. FIG. 18A shows AtCERK1 bindingto chitopentaose (Chitin (CO5)). FIG. 18B shows AtCERK1 binding tochitooctaose (Chitin (CO8)). For FIGS. 18A-18B, seven 2-fold dilutionseries of analyte (1.56-100 μM) were used for each experiment;experimental binding curves are represented in solid lines, fittingcurves in dashed lines; goodness of fit is described by the global fitR² on the mean value of each point; number of replicates performed usingindependent protein preparations (n) indicated; and kinetic parameters(k_(on) and k_(off)) are shown. FIG. 18C shows a model of CO perceptionby CO receptors (e.g., CERK1, LYK5). FIG. 18D shows a model of LCOperception by LCO receptors (e.g., NFP, LYK). FIG. 18E shows a model ofLCO perception by hydrophobic patch mutant LCO receptors (e.g., NFP,LYK). FIG. 18F shows structural alignment of the ectodomains of MedicagoNFP, Arabidopsis CERK1 and Lotus CERK6. Molecular fits (RMSD values)based on structural superposition of the ectodomains are shown in Å(Angstrom). The structures (above) are shaded according to the schematicrepresentation of the ectodomain (below). The conserved disulfideconnectivity pattern between Medicago NFP, Arabidopsis CERK1 and LotusCERK6 is highlighted.

FIGS. 19A-19D show BLI binding curves for WT NFP-ECD and hydrophobicpatch mutant NFP-ECD (L147D/L154D) binding to S. meliloti LCO-IV and aschematic of the NFP receptor. FIG. 19A shows WT NFP-ECD binding to S.meliloti LCO-IV. FIG. 19B shows L147D/L154D NFP-ECD binding to S.meliloti LCO-IV. For FIGS. 19A-19B, seven 2-fold dilution series ofanalyte (1.56-100 μM) were used for each experiment; and experimentalbinding curves are represented in solid lines, fitting curves in dashedlines. FIG. 19C shows a table summarizing the kinetic parameters ofFIGS. 19A-19B, with goodness of fit described by the global fit R² onthe mean value of each point, and number of replicates performed usingindependent protein preparations (n) indicated. FIG. 19D shows aschematic of the NFP receptor with LysM1, LysM2, LysM3, stem, andtransmembrane (TM) and kinase domains labeled, and the location of thehydrophobic patch in LysM2 indicated by a grey bar. Numbers below theschematic provide the corresponding amino acid residues, and thelocations of the CxC motifs flanking the LysM domains are shown.

DETAILED DESCRIPTION

The following description sets forth exemplary methods, parameters, andthe like. It should be recognized, however, that such description is notintended as a limitation on the scope of the present disclosure but isinstead provided as a description of exemplary embodiments.

Modified Plant LysM Receptors

Certain aspects of the present disclosure relate to a modified plantLysM receptor comprising a LysM2 domain modified to comprise ahydrophobic patch on the surface of the LysM2 domain. In someembodiments, the modified LysM2 domain binds a lipo-chitooligosaccharide(LCO). In some embodiments, the modified LysM2 domain binds the LCO withhigher affinity than the unmodified LysM2 domain. In some embodiments,the modified LysM2 domain binds the LCO with higher selectivity for theLCO than the unmodified LysM2 domain. In some embodiments, the higheraffinity or higher selectivity is due to the hydrophobic patchinteracting with the LCO. In some embodiments, the higher affinity orhigher selectivity is due to the hydrophobic patch interacting with thelipid of the LCO. In some embodiments, the LCO is produced bynitrogen-fixing bacteria or by mycorrhizal fungi. In some embodiments,the LCO is produced by nitrogen-fixing bacteria selected from the groupof Mesorhizobium loti, Mesorhizobium huakuii, Mesorhizobiummediterraneum, Mesorhizobium ciceri, Mesorhizobium spp., Rhizobiummongolense, Rhizobium tropici, Rhizobium etli phaseoli, Rhizobiumgiardinii, Rhizobium leguminosarum optionally R. leguminosarum trifolii,R. leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., and any combination thereof,or by mycorrhizal fungi selected from the group consisting ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, or any combination thereof.

In some embodiments of any of the above embodiments, the LysM receptoris selected from the group consisting of a LysM chitooligosaccharide(CO) receptor, a LysM LCO receptor, and a LysM peptidoglycan (PGN)receptor. In some embodiments, the hydrophobic patch is adjacent to achitin binding motif if the LysM receptor is the LysM CO receptor or theLysM LCO receptor. In some embodiments, the hydrophobic patch is within30 Å, 29 Å, 28 Å, 27 Å, 26 Å, 25 Å, 24 Å, 23 Å, 22 Å, 21 Å, 20 Å, 19 Å,18 Å, 17 Å, 16 Å, 15 Å, 14 Å, 13 Å, 12 Å, 11 Å, 10 Å, 9.5 Å, 9 Å, 8.5 Å,8 Å, 7.5 Å, 7 Å, 6.5 Å, 6 Å, 5.5 Å, 5 Å, 4.5 Å, 4 Å, 3.5 Å, 3 Å, 2.5 Å,2 Å, 1.5 Å, or 1 Å of a chitin binding motif if the LysM receptor is theLysM CO receptor or the LysM LCO receptor. In some embodiments, thehydrophobic patch is adjacent to a glycan binding motif if the LysMreceptor is the LysM PGN receptor. In some embodiments, the hydrophobicpatch is within 30 Å, 29 Å, 28 Å, 27 Å, 26 Å, 25 Å, 24 Å, 23 Å, 22 Å, 21Å, 20 Å, 19 Å, 18 Å, 17 Å, 16 Å, 15 Å, 14 Å, 13 Å, 12 Å, 11 Å, 10 Å, 9.5Å, 9 Å, 8.5 Å, 8 Å, 7.5 Å, 7 Å, 6.5 Å, 6 Å, 5.5 Å, 5 Å, 4.5 Å, 4 Å, 3.5Å, 3 Å, 2.5 Å, 2 Å, 1.5 Å, or 1 Å of a glycan binding motif if the LysMreceptor is the LysM PGN receptor. In some embodiments, the LysMreceptor is not an exopolysaccharide (EPS) receptor.

In some embodiments of any of the above embodiments, the LysM receptoris a polypeptide with at least 70% sequence identity, at least 71%, 72%,73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least99% sequence identity to SEQ ID NO:34 (i.e., Lotus CERK6;BAI79273.1_LjCERK6). In some embodiments of any of the aboveembodiments, the LysM receptor is a polypeptide having amino acidsequence SEQ ID NO:34 (i.e., Lotus CERK6; BAI79273.1_LJCERK6).

In some embodiments of any of the above embodiments, the hydrophobicpatch was generated by deleting at least one non-hydrophobic amino acidresidue, substituting at least one amino acid residue with a morehydrophobic amino acid, or combinations thereof. In some embodiments,the at least one amino acid was identified by an amino acid sequencealignment with a LysM2 domain from a LysM high affinity LCO receptorthat naturally has a hydrophobic patch that interacts with LCO. In someembodiments, the at least one amino acid corresponds to an amino acidthat is in bold in FIGS. 12A-12G and FIG. 13C. In some embodiments, theat least one amino acid corresponds to an amino acid that is in bold inFIGS. 12A-12G and FIG. 13C or corresponds to an amino acid that isimmediately N-terminal or C-terminal to an amino acid that is in bold inFIGS. 12A-12G and FIG. 13C. In some embodiments, the at least one aminoacid corresponds to an amino acid that is in bold in FIGS. 12A-12G in aknown LCO receptor. In some embodiments, the at least one amino acidcorresponds to an amino acid that is in bold in FIGS. 12A-12G in a knownLCO receptor or corresponds to an amino acid that is immediatelyN-terminal or C-terminal to an amino acid that is in bold in FIGS.12A-12G in a known LCO receptor. In some embodiments, the at least oneamino acid was identified by structural modelling to identify a regionin LysM2 where the hydrophobic patch can be engineered. In someembodiments, the structural modeling used the unmodified plant LysMamino acid sequence and a LysM domain three dimensional structure thathas a known hydrophobic patch. In some embodiments, the LysM domainthree dimensional structure is a Medicago NFP ectodomain. In someembodiments, the known hydrophobic patch amino acid residues of the LysMdomain three dimensional structure are or correspond to L147, L151,L152, L154, T156, K157 and V158 of the Medicago NFP ectodomain. In someembodiments, the alpha carbon of at least one amino acid was within 3 Åof an alpha carbon of a known hydrophobic patch amino acid residue inthe structural alignment. In some embodiments, the structural modelingwas performed using SWISS-MODEL, PDB2PQR, APBS, PyMol, and APBS tools2.1.

In some aspects, the present disclosure relates to a modified plant LysMreceptor comprising a first LysM1 domain modified to replace at leastpart of the first LysM1 domain with at least part of a second LysM1domain. In some embodiments, the first LysM1 domain is modified bysubstituting a first part of the first LysM1 domain with a third part ofa second LysM1 domain and/or by substituting a second part of the firstLysM1 domain with a fourth part of the second LysM1 domain. In someembodiments, the first LysM1 domain and the second LysM1 domain havedifferent affinities and/or selectivities for oligosaccharides and themodification of the first LysM1 domain alters the affinity, selectivity,and/or specificity to be more like the second LysM1 domain. In someembodiments, the first part and the third part correspond to SEQ IDNO:30 [Lotus CERK6 region II 43-53] or NGSNLTYISEI, SEQ ID NO:28 [LotusNFR1 region II 41-52] or PGVFILQNITTF; and wherein the second part andthe fourth part correspond to SEQ ID NO:31 [Lotus CERK6 region IV 74-82]or ASKDSVQAG; SEQ ID NO:29 [Lotus NFR1 region IV 73-81], or LNDINIQSF.In some embodiments, the first LysM1 domain is selected from the groupof SEQ ID NO:32 [LysM1 domain Lotus NFR1; LjNFR1/26-95], SEQ ID NO:33[LysM1 domain Medicago LYK3; MtLYK3/25-95], or NFR1DLALASYYILPGVFILQNITTFMQSEIVSSNDAITSYNKDKILNDINIQSFQRLNIPFP; and thesecond LysM1 domain is CERK6:ALAQASYYLLNGSNLTYISEIMQSSLLTKPEDIVSYNQDTIASKDSVQAGQRINVPFP. In someembodiments, the first part is selected from SEQ ID NO:30 [Lotus CERK6region II 43-53] or NGSNLTYISEI; the second part is selected from SEQ IDNO:31 [Lotus CERK6 region IV 74-82] or ASKDSVQAG; the third part isselected from SEQ ID NO:28 [Lotus NFR1 region II 41-52] or PGVFILQNITTF;and the fourth part is selected from SEQ ID NO:29 [Lotus NFR1 region IV73-81] or LNDINIQSF. In some embodiments, the entire first LysM1 domainwas replaced with the entire second LysM1 domain. In some embodiments,the modified LysM1 domain binds a lipo-chitooligosaccharide (LCO)produced by nitrogen-fixing bacteria or by mycorrhizal fungi. In someembodiments, the LCO is produced by nitrogen-fixing bacteria selectedfrom the group consisting of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., and any combinationthereof, or by mycorrhizal fungi selected from the group consisting ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, and any combination thereof. Insome embodiments, the modified LysM1 domain binds an LCO with higheraffinity than an unmodified LysM1 domain. In some embodiments, themodified LysM1 domain binds LCOs with higher selectivity than anunmodified LysM1 domain. In some embodiments, the modified LysM1 domainbinds LCOs with altered specificity as compared to an unmodified LysM1domain. In some embodiments, structural modelling was used to define theLysM1 domain and was used to identify the first part, the second part,the third part, and/or the fourth part for substitution. In someembodiments, the receptor of the above embodiments further contains aLysM2 domain modified to contain a hydrophobic patch as in any one ofthe previous embodiments relating to modifications to the LysM2 domain.

Additional aspects of the present disclosure relate to a modified plantLysM receptor including a LysM2 domain modified to include a hydrophobicpatch on the surface of the LysM2 domain. In some embodiments, themodified LysM2 domain binds a lipo-chitooligosaccharide (LCO). In someembodiments, the modified LysM2 domain binds the LCO with higheraffinity than the unmodified LysM2 domain. In some embodiments, themodified LysM2 domain binds the LCO with higher selectivity for the LCOthan the unmodified LysM2 domain. In some embodiments, the higheraffinity or higher selectivity is due to the hydrophobic patchinteracting with the LCO. In some embodiments, the higher affinity orhigher selectivity is due to the hydrophobic patch interacting with thelipid of the LCO. In some embodiments, the LCO is produced bynitrogen-fixing bacteria or by mycorrhizal fungi. In some embodiments,the LCO is produced by nitrogen-fixing bacteria selected from the groupof Mesorhizobium loti, Mesorhizobium huakuii, Mesorhizobiummediterraneum, Mesorhizobium ciceri, Mesorhizobium spp., Rhizobiummongolense, Rhizobium tropici, Rhizobium etli phaseoli, Rhizobiumgiardinii, Rhizobium leguminosarum optionally R. leguminosarum trifolii,R. leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., or any combination thereof,or by mycorrhizal fungi selected from the group of Acaulosporaceae spp.,Diversisporaceae spp., Gigasporaceae spp., Pacisporaceae spp.,Funneliformis spp., Glomus spp., Rhizophagus spp., Sclerocystis spp.,Septoglomus spp., Claroideoglomus spp., Ambispora spp., Archaeosporaspp., Geosiphon pyriformis, Paraglomus spp., other species in thedivision Glomeromycota, or any combination thereof.

In some embodiments of any of the above embodiments, the LysM receptoris selected from the group of a LysM chitooligosaccharide (CO) receptor,a LysM LCO receptor, or a LysM peptidoglycan (PGN) receptor. In someembodiments, the hydrophobic patch is adjacent to a chitin binding motifif the LysM receptor is the LysM CO receptor or the LysM LCO receptor.In some embodiments, the hydrophobic patch is within 30 Å, 29 Å, 28 Å,27 Å, 26 Å, 25 Å, 24 Å, 23 Å, 22 Å, 21 Å, 20 Å, 19 Å, 18 Å, 17 Å, 16 Å,15 Å, 14 Å, 13 Å, 12 Å, 11 Å, 10 Å, 9.5 Å, 9 Å, 8.5 Å, 8 Å, 7.5 Å, 7 Å,6.5 Å, 6 Å, 5.5 Å, 5 Å, 4.5 Å, 4 Å, 3.5 Å, 3 Å, 2.5 Å, 2 Å, 1.5 Å, or 1Å of a chitin binding motif if the LysM receptor is the LysM CO receptoror the LysM LCO receptor. In some embodiments, the hydrophobic patch isadjacent to a glycan binding motif if the LysM receptor is the LysM PGNreceptor. In some embodiments, the hydrophobic patch is within 30 Å, 29Å, 28 Å, 27 Å, 26 Å, 25 Å, 24 Å, 23 Å, 22 Å, 21 Å, 20 Å, 19 Å, 18 Å, 17Å, 16 Å, 15 Å, 14 Å, 13 Å, 12 Å, 11 Å, 10 Å, 9.5 Å, 9 Å, 8.5 Å, 8 Å, 7.5Å, 7 Å, 6.5 Å, 6 Å, 5.5 Å, 5 Å, 4.5 Å, 4 Å, 3.5 Å, 3 Å, 2.5 Å, 2 Å, 1.5Å, or 1Λ of a glycan binding motif if the LysM receptor is the LysM PGNreceptor. In some embodiments, the LysM receptor is not anexopolysaccharide (EPS) receptor.

In some embodiments of any of the above embodiments, the LysM receptoris a polypeptide having at least 70% sequence identity, at least 71%,72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or atleast 99% sequence identity to SEQ ID NO:34 (i.e., Lotus CERK6;BAI79273.1_LjCERK6). In some embodiments of any of the aboveembodiments, the LysM receptor is a polypeptide having amino acidsequence SEQ ID NO:34 (i.e., Lotus CERK6; BAI79273.1_LjCERK6).

In some embodiments of any of the above embodiments, the hydrophobicpatch was generated by deleting at least one non-hydrophobic amino acidresidue, substituting at least one amino acid residue with a morehydrophobic amino acid, or combinations thereof. In some embodiments ofany of the above embodiments, the hydrophobic patch was generated bymodifying an existing hydrophobic patch in the unmodified LysM receptor.In some embodiments, the unmodified LysM receptor was modified bydeleting at least one non-hydrophobic amino acid residue, substitutingat least one amino acid residue with a more hydrophobic amino acid,substituting at least one hydrophobic amino acid residue with anotherhydrophobic amino acid residue, or combinations thereof. In someembodiments, the at least one amino acid was identified by an amino acidsequence alignment with a LysM2 domain from a LysM high affinity LCOreceptor that naturally has a hydrophobic patch that interacts with LCO.In some embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G and FIG. 13C. In someembodiments, the at least one amino acid corresponds to an amino acidthat is in bold underline in FIGS. 12A-12G and FIG. 13C or correspondsto an amino acid that is immediately N-terminal or C-terminal to anamino acid that is in bold underline in FIGS. 12A-12G and FIG. 13C. Insome embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G in a known LCO receptor.In some embodiments, the at least one amino acid corresponds to an aminoacid that is in bold underline in FIGS. 12A-12G in a known LCO receptoror corresponds to an amino acid that is immediately N-terminal orC-terminal to an amino acid that is in bold underline in FIGS. 12A-12Gin a known LCO receptor. In some embodiments, the at least one aminoacid was identified by structural modelling to identify a region inLysM2 where the hydrophobic patch can be engineered. In someembodiments, the structural modeling used the unmodified plant LysMamino acid sequence and a LysM domain three dimensional structure thathas a known hydrophobic patch. In some embodiments, the LysM domainthree dimensional structure is a Medicago NFP ectodomain. In someembodiments, the known hydrophobic patch amino acid residues of the LysMdomain three dimensional structure are or correspond to L147, L151,L152, L154, T156, K157 and V158 of the Medicago NFP ectodomain. In someembodiments, the LysM domain three dimensional structure is a LotusLYS11 ectodomain. In some embodiments, the unmodified LysM receptor isthe Lotus LYS11 receptor and the existing hydrophobic patch amino acidresidues of the LysM domain that are modified are or correspond to K100,E101, G102, E103, S104, Y105, Y106, N128, and Y129 of the Lotus LYS11ectodomain. In some embodiments, the alpha carbon of at least one aminoacid was within 3 Å of an alpha carbon of a known hydrophobic patchamino acid residue in the structural alignment. In some embodiments, thestructural modeling was performed using SWISS-MODEL, PDB2PQR, APBS,PyMol, and APBS tools 2.1. In some embodiments of any of the aboveembodiments, either or both (i) 80% or fewer, 79% or fewer, 78% orfewer, 77% or fewer, 76% or fewer, 75% or fewer, 74% or fewer, 73% orfewer, 72% or fewer, 71% or fewer, 70% or fewer, 69% or fewer, 68% orfewer, 67% or fewer, 66% or fewer, 65% or fewer, 64% or fewer, 63% orfewer, 62% or fewer, 61% or fewer, 60% or fewer, 59% or fewer, 58% orfewer, 57% or fewer, 56% or fewer, 55% or fewer, 54% or fewer, 53% orfewer, 52% or fewer, 51% or fewer, 50% or fewer, 49% or fewer, 48% orfewer, 47% or fewer, 46% or fewer, 45% or fewer, 44% or fewer, 43% orfewer, 42% or fewer, 41% or fewer, 40% or fewer, 39% or fewer, 38% orfewer, 37% or fewer, 36% or fewer, 35% or fewer, 34% or fewer, 33% orfewer, 32% or fewer, 31% or fewer, 30% or fewer, 29% or fewer, 28% orfewer, 27% or fewer, 26% or fewer, 25% or fewer, 24% or fewer, 23% orfewer, 22% or fewer, 21% or fewer, or 20% or fewer of amino acidresidues in the LysM2 domain of the unmodified LysM receptor weresubstituted or deleted to generate the modified plant LysM receptor, and(ii) the entire LysM2 domain in the unmodified plant LysM receptor wasnot substituted with another entire LysM2 domain to generate themodified plant LysM receptor. In some embodiments of any of the aboveembodiments, the unmodified plant LysM receptor was selected using themethod of any one of the aspects of the present disclosure relating tosuch selection including any and all embodiments thereof.

In some aspects, the present disclosure relates to a modified plant LysMreceptor including a first LysM1 domain modified to replace at leastpart of the first LysM1 domain with at least part of a second LysM1domain. In some embodiments, the first LysM1 domain is modified bysubstituting a first part of the first LysM1 domain with a third part ofa second LysM1 domain and/or by substituting a second part of the firstLysM1 domain with a fourth part of the second LysM1 domain. In someembodiments, the first LysM1 domain and the second LysM1 domain havedifferent affinities, selectivities, and/or specificities foroligosaccharides and the modification of the first LysM1 domain altersthe affinity, selectivity, and/or specificity to be more like the secondLysM1 domain. In some embodiments, the first part and the third partcorrespond to SEQ ID NO:30 [Lotus CERK6 region II 43-53] or NGSNLTYISEI,SEQ ID NO:28 [Lotus NFR1 region II 41-52] or PGVFILQNITTF; and whereinthe second part and the fourth part correspond to SEQ ID NO:31 [LotusCERK6 region IV 74-82] or ASKDSVQAG; SEQ ID NO:29 [Lotus NFR1 region IV73-81], or LNDINIQSF. In some embodiments, the first LysM1 domain isselected from the group of SEQ ID NO:32 [LysM1 domain Lotus NFR1;LjNFR1/26-95], SEQ ID NO:33 [LysM1 domain Medicago LYK3; MtLYK3/25-95],or NFR1 DLALASYYILPGVFILQNITTFMQSEIVSSNDAITSYNKDKILNDINIQSFQRLNIPFP; andthe second LysM1 domain is CERK6:ALAQASYYLLNGSNLTYISEIMQSSLLTKPEDIVSYNQDTIASKDSVQAGQRINVPFP. In someembodiments, the first part is selected from SEQ ID NO:30 [Lotus CERK6region II 43-53] or NGSNLTYISEI; the second part is selected from SEQ IDNO:28 [Lotus CERK6 region IV 74-82] or ASKDSVQAG; the third part isselected from SEQ ID NO:31 [Lotus NFR1 region II 41-52] or PGVFILQNITTF;and the fourth part is selected from SEQ ID NO:29 [Lotus NFR1 region IV73-81] or LNDINIQSF. In some embodiments of any of the above embodimentsincluding the first LysM1 domain being modified to replace at least partof the first LysM1 domain with at least part of a second LysM1 domain,the first LysM1 domain is further modified by substituting a fifth partof the first LysM1 domain with a sixth part of a second LysM1 domain. Insome embodiments, the first LysM1 domain is SEQ ID NO:115 [LysM1 domainLotus NFR1; LjNFR1/32-89] or SEQ ID NO:106 [LysM1 domain Lotus NFR1;LjNFR1/31-89] and the second LysM1 domain is SEQ ID NO:114 [LysM1 domainMedicago LYK3; MtLYK3/31-89] or SEQ ID NO:105 [LysM1 domain MedicagoLYK3; MtLYK3/30-89]. In some embodiments, wherein the fifth part is SEQID NO:53 [Lotus NFR1 region III 59-62; LjNFR1/56-92], and wherein thesixth part is SEQ ID NO:46 [Medicago LYK3 region III 57-62;MtLYK3/57-62]. In some embodiments, the first LysM1 domain is modifiedby substituting a seventh part of the first LysM1 domain, wherein theseventh part spans the first part of the first LysM1 domain, the secondpart of the first LysM1 domain, and the fifth part of the first LysM1domain, with an eighth part of the second LysM1 domain, wherein theeighth part spans the third part of the second LysM1 domain, the fourthpart of the second LysM1 domain, and the sixth part of the second LysM1domain. In some embodiments, the seventh part of the first LysM1 domainis SEQ ID NO:51 [Lotus NFR1 regions II-IV 41-82; LjNFR1/41-82], and theeighth part of the second LysM1 domain is SEQ ID NO:113 [Medicago LYK3regions II-IV 40-82; MtLYK3/40-82] or SEQ ID NO:104 [Medicago LYK3regions II-IV 41-82; MtLYK3/41-82]. In some embodiments, the first LysM1domain is SEQ ID NO:33 [LysM1 domain Medicago LYK3; MtLYK3/31-89] andthe second LysM1 domain is SEQ ID NO:32 [LysM1 domain Lotus NFR1;LjNFR1/32-89]. In some embodiments, the fifth part is SEQ ID NO:46[Medicago LYK3 region III 57-62; MtLYK3/57-62], and the sixth part isSEQ ID NO:53 [Lotus NFR1 region III 59-62; LjNFR1/59-62]. In someembodiments of any of the above embodiments including the first LysM1domain being SEQ ID NO:33 and the second LysM1 domain being SEQ IDNO:32, the first LysM1 domain is modified by substituting a seventh partof the first LysM1 domain, wherein the seventh part spans the first partof the first LysM1 domain, the second part of the first LysM1 domain,and the fifth part of the first LysM1 domain, with an eighth part of thesecond LysM1 domain, wherein the eighth part spans the third part of thesecond LysM1 domain, the fourth part of the second LysM1 domain, and thesixth part of the second LysM1 domain. In some embodiments, the seventhpart of the first LysM1 domain is SEQ ID NO:51 [Lotus NFR1 regions II-IV41-82; LjNFR1/41-82], and the eighth part of the second LysM1 domain isSEQ ID NO:113 [Medicago LYK3 regions II-IV 40-82; MtLYK3/40-82] or SEQID NO:104 [Medicago LYK3 regions II-IV 41-82; MtLYK3/41-82].

In some embodiments of any of the above embodiments including the firstLysM1 domain being modified to replace at least part of the first LysM1domain with at least part of a second LysM1 domain, the entire firstLysM1 domain was replaced with the entire second LysM1 domain. In someembodiments of any of the above embodiments including the first LysM1domain being modified to replace at least part of the first LysM1 domainwith at least part of a second LysM1 domain, either or both (i) 80% orfewer, 79% or fewer, 78% or fewer, 77% or fewer, 76% or fewer, 75% orfewer, 74% or fewer, 73% or fewer, 72% or fewer, 71% or fewer, 70% orfewer, 69% or fewer, 68% or fewer, 67% or fewer, 66% or fewer, 65% orfewer, 64% or fewer, 63% or fewer, 62% or fewer, 61% or fewer, 60% orfewer, 59% or fewer, 58% or fewer, 57% or fewer, 56% or fewer, 55% orfewer, 54% or fewer, 53% or fewer, 52% or fewer, 51% or fewer, 50% orfewer, 49% or fewer, 48% or fewer, 47% or fewer, 46% or fewer, 45% orfewer, 44% or fewer, 43% or fewer, 42% or fewer, 41% or fewer, 40% orfewer, 39% or fewer, 38% or fewer, 37% or fewer, 36% or fewer, 35% orfewer, 34% or fewer, 33% or fewer, 32% or fewer, 31% or fewer, 30% orfewer, 29% or fewer, 28% or fewer, 27% or fewer, 26% or fewer, 25% orfewer, 24% or fewer, 23% or fewer, 22% or fewer, 21% or fewer, or 20% orfewer of amino acid residues in the first LysM1 domain were substitutedor deleted with the corresponding amino acid residues of the secondLysM1 domain, and (ii) the entire LysM1 domain in the unmodified plantLysM receptor was not substituted with another entire LysM2 domain togenerate the modified plant LysM receptor. In some embodiments, themodified LysM1 domain binds a lipo-chitooligosaccharide (LCO) producedby nitrogen-fixing bacteria or by mycorrhizal fungi. In someembodiments, the LCO is produced by nitrogen-fixing bacteria selectedfrom the group of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., or any combinationthereof, or by mycorrhizal fungi selected from the group ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, or any combination thereof. Insome embodiments, the modified LysM1 domain binds an LCO with higheraffinity than an unmodified LysM1 domain. In some embodiments, themodified LysM1 domain binds LCOs with higher selectivity than anunmodified LysM1 domain. In some embodiments, the modified LysM1 domainbinds LCOs with altered specificity as compared to an unmodified LysM1domain. In some embodiments, structural modelling was used to define theLysM1 domain and was used to identify the first part, the second part,the third part, and/or the fourth part for substitution. In someembodiments, the unmodified plant LysM receptor was selected using themethod of any one of the aspects of the present disclosure relating tosuch selection including any and all embodiments thereof and the secondLysM2 domain is from the donor plant LysM receptor. In some embodiments,the receptor of the above embodiments further contains a LysM2 domainmodified to contain a hydrophobic patch as in any one of the previousembodiments relating to modifying the LysM2 domain.

LysM receptors are a well known and well understood type of receptor.LysM receptors have three characteristic domains located in theectodomain of the protein: LysM1, LysM2, and LysM3, which are present inthis order on the protein sequence. The LysM1 domain is located towardthe N-terminal end of the protein sequence, and is preceded by anN-terminal signal peptide as well as a C(x)xxxC motif. The LysM1 domainis separated from the LysM2 domain by a CxC motif, and the LysM2 domainis separated from the LysM3 domain by a CxC motif as well. The threeLysM domains, as well as the C(x)xxxC and CxC motif are clearly shown inFIGS. 8A-8C, FIGS. 9A-9B, FIGS. 10A-10B, and FIGS. 11A-11B that showindividual alignments of Nod factor (e.g., LCO) LysM receptors, EPS LysMreceptors, and chitin (CO) as well as PGN LysM receptors, again clearlydepicting the three LysM domains as well as the C(x)xxxC and CxC motifs.The category of LysM receptors is therefore known by one of skill in theart.

As used in the present disclosure, the term “affinity” refers toaffinity for LCOs generally. The LysM receptors of the presentdisclosure may contain a hydrophobic patch in their LysM2 domain.Without wanting to be limited to theory, it is believed that LysMreceptors with the hydrophobic patch have higher affinity for LCOs ascompared to LysM receptors without the hydrophobic patch, but LysMreceptors with domain-swapped LysM1 domains would also provide higheraffinity for LCOs and other agonists. Affinity can be measured using themethods described in the Examples below, and using other methods knownin the art that measure binding kinetics, association, dissociation, andK_(D).

As used in the present disclosure, the term “selectivity” refers to thedifferentiation between different polysaccharide ligands, specificallybetween lipo-chitooligosaccharides (LCOs) as a class and otherpolysaccharide ligands, preferably chitooligosaccharides (COs). Withoutwanting to be limited to theory, it is believed that this hydrophobicpatch confers selective recognition of LCOs over COs, and that thereforeLysM receptors with the hydrophobic patch have increased selectivity ascompared to LysM receptors without the hydrophobic patch. In addition,the LysM receptors with domain-swapped LysM1 domains should also havehigher or altered selectivity depend upon the choice of the donorreceptor.

As used in the present disclosure, the term “specificity” refers to thedifferentiation between different lipo-chitooligosaccharides (LCOs)produced by different nitrogen-fixing bacterial species and/ormycorrhizal fungi. The LysM receptors of the present disclosure maycontain a LysM1 domain where regions (e.g., partial, entire) in theLysM1 domain have been replaced with the corresponding regions of theLysM1 domain from a donor LysM receptor. Without wanting to be limitedto theory, it is believed that if the donor LysM receptor is a highaffinity and specificity LCO LysM receptor such as a legume NFR1receptor, this replacement can alter the specificity of the LysMreceptor, but LysM receptors with a hydrophobic patch in the LysM2domain may also provide specificity for specific LCOs. The LysM1 domainis clearly shown in FIG. 14, which shows an alignment between Lotus NFR1and Lotus CERK6, and clearly designates region II and region IV withinthe LysM1 domain. LysM1 domain replacement can confer highly specificrecognition of LCOs produced by particular nitrogen-fixing bacterialspecies and/or mycorrhizal fungal species, and therefore LysM receptorswith the replaced domain can have altered specificity as compared toLysM receptors without the replaced domain, which allows the modifiedreceptors to recognize different nitrogen-fixing bacterial speciesand/or mycorrhizal fungal species. For at least these reasons, the highaffinity, high selectivity, and/or high specificity LysM receptors ofthe present disclosure will be readily understood by one of skill in theart.

Genetically Altered Plants and Seeds

In some aspects, the present disclosure relates to a genetically alteredplant or part thereof, comprising a nucleic acid sequence encoding amodified plant LysM receptor of any one of the embodiments described inthe section “Modified plant LysM receptors”. In some embodiments, themodified plant LysM receptor has higher selectivity and/or affinity forLCOs than the unmodified plant LysM receptor and the expression of themodified plant LysM receptor allows the plant or part thereof torecognize LCOs with high selectivity and/or affinity. In someembodiments, the LCOs are produced by nitrogen-fixing bacteria or bymycorrhizal fungi. In some embodiments, the LCOs are produced bynitrogen-fixing bacteria selected from the group consisting ofMesorhizobium loti, Mesorhizobium huakuii, Mesorhizobium mediterraneum,Mesorhizobium ciceri, Mesorhizobium spp., Rhizobium mongolense,Rhizobium tropici, Rhizobium etli phaseoli, Rhizobium giardinii,Rhizobium leguminosarum optionally R. leguminosarum trifolii, R.leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., and any combination thereof,or by or by mycorrhizal fungi selected from the group consisting ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, and any combination thereof. Insome embodiments, the modified polypeptide is localized to a plant cellplasma membrane. In some embodiments, the plant cell is a root cell. Insome embodiments, the root cell is a root epidermal cell. In someembodiments, the root cell is a root cortex cell. In some embodiments,the modified polypeptide is expressed in a developing plant root system.In some embodiments, the nucleic acid sequence is operably linked to apromoter. In some embodiments, the promoter is a root specific promoter.In some embodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO:24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter, a derivative of the CaMV35Spromoter, the maize ubiquitin promoter, the trefoil promoter, a veinmosaic cassava virus promoter, or the Arabidopsis UBQ10 promoter. Insome embodiments, the plant is selected from the group of corn (e.g.,maize, Zea mays), rice (e.g., Oryza sativa, Oryza glaberrima, Zizaniaspp.), barley (e.g., Hordeum vulgare), wheat (e.g., common wheat, spelt,durum, Triticum aestivum, Triticum spelta, Triticum durum, Triticumspp.), Trema spp. (e.g., Trema cannabina, Trema cubense, Trema discolor,Trema domingensis, Trema integerrima, Trema lamarckiana, Tremamicrantha, Trema orientalis, Trema philippinensis, Trema strigilosa,Trema tomentosa), apple (e.g., Malus pumila), pear (e.g., Pyruscommunis, Pyrus×bretschneideri, Pyrus pyrifolia, Pyrus sinkiangensis,Pyrus pashia, Pyrus spp.), plum (e.g., prune, damson, bullaces, Prunusdomestica, Prunus salicina), apricot (e.g., Prunus armeniaca, Prunusbrigantina, Prunus mandshurica, Prunus mume, Prunus sibirica), peach(e.g., nectarine, Prunus persica), almond (e.g., Prunus dulcis, Prunusamygdalus), walnut (e.g., Persian walnut, English walnut, black walnut,Juglans regia, Juglans nigra, Juglans cinerea, Juglans californica),strawberry (e.g., Fragaria×ananassa, Fragaria chiloensis, Fragariavirginiana, Fragaria vesca), raspberry (e.g., European red raspberry,black raspberry, Rubus idaeus, Rubus occidentalis, Rubus strigosus),blackberry (e.g., evergreen blackberry, Himalayan blackberry, Rubusfruticosus, Rubus ursinus, Rubus laciniatus, Rubus argutus, Rubusarmeniacus, Rubus plicatus, Rubus ulmifolius, Rubus allegheniensis), redcurrant (e.g., Ribes rubrum, Ribes spicatum, Ribesbes alpinum, Ribesschlechtendalii, Ribes multiflorum, Ribes petraeum, Ribes triste), blackcurrant (e.g., Ribes nigrum), melon (e.g., watermelon, winter melon,casabas, cantaloupe, honeydew, muskmelon, Citrullus lanatus, Benincasahispida, Cucumis melo cantalupensis, Cucumis melo inodorus, Cucumis meloreticulatus), cucumber (e.g., slicing cucumbers, pickling cucumbers,English cucumber, Cucumis sativus), pumpkin (e.g., Cucurbita pepo,Cucurbita maxima), squash (e.g., gourd, Cucurbita argyrosperma,Cucurbita ficifolia, Cucurbita maxima, Cucurbita moschata), grape (e.g.,Vitis vinifera, Vitis amurensis, Vitis labrusca, Vitis mustangensis,Vitis riparia, Vitis rotundifolia), bean (e.g., Phaseolus vulgaris,Phaseolus lunatus, Vigna angularis, Vigna radiate, Vigna mungo,Phaseolus coccineus, Vigna umbellate, Vigna acontifolia, Phaseolusacutifolius, Vicia faba, Vicia faba equine, Phaseolus spp., Vigna spp.),soybean (e.g., soy, soya bean, Glycine max, Glycine soja), pea (e.g.,Pisum spp., Pisum sativum var. sativum, Pisum sativum var. arvense),chickpea (e.g., garbanzo, Bengal gram, Cicer arietinum), cowpea (e.g.,black-eyed pea, blackeye bean, Vigna unguiculata), pigeon pea (e.g.,Arhar/Toor, cajan pea, Congo bean, gandules, Caganus cajan), lentil(e.g., Lens culinaris), Bambara groundnut (e.g., earth pea, Vignasubterranea), lupin (e.g., Lupinus spp.), pulses (e.g., minor pulses,Lablab purpureaus, Canavalia ensiformis, Canavalia gladiate,Psophocarpus tetragonolobus, Mucuna pruriens var. utilis, Pachyrhizuserosus), Medicago spp. (e.g., Medicago sativa, Medicago truncatula,Medicago arborea), Lotus spp. (e.g., Lotus japonicus), forage legumes(e.g., Leucaena spp., Albizia spp., Cyamopsis spp., Sesbania spp.,Stylosanthes spp., Trifolium spp., Vicia spp.), indigo (e.g., Indigoferaspp., Indigofera tinctoria, Indigofera suffruticosa, Indigoferaarticulata, Indigofera oblongifolia, Indigofera aspalthoides, Indigoferasuffruticosa, Indigofera arrecta), legume trees (e.g., locust trees,Gleditsia spp., Robinia spp., Kentucky coffeetree, Gymnocladus dioicus,Acacia spp., Laburnum spp., Wisteria spp.), or hemp (e.g., cannabis,Cannabis sativa).

In some aspects, the present disclosure relates to a genetically alteredplant or part thereof, comprising a first nucleic acid sequence encodinga modified plant LysM receptor where the LysM1 domain has been modifiedas in any of the preceding embodiments relating to modification of theLysM1 domain and a second nucleic acid sequence encoding a modifiedplant LysM receptor where the LysM2 domain has been modified to includea hydrophobic patch as in any of the preceding embodiments relating tomodification of the LysM2 domain. In some embodiments, the modifiedplant LysM receptor has higher selectivity and/or affinity for LCOs thanthe unmodified plant LysM receptor and the expression of the modifiedplant LysM receptor allows the plant or part thereof to recognize LCOswith high selectivity and/or affinity. In some embodiments, the LCOs areproduced by nitrogen-fixing bacteria or by mycorrhizal fungi. In someembodiments, the LCOs are produced by nitrogen-fixing bacteria selectedfrom the group consisting of Mesorhizobium loti, Mesorhizobium huakuii,Mesorhizobium mediterraneum, Mesorhizobium ciceri, Mesorhizobium spp.,Rhizobium mongolense, Rhizobium tropici, Rhizobium etli phaseoli,Rhizobium giardinii, Rhizobium leguminosarum optionally R. leguminosarumtrifolii, R. leguminosarum viciae, and R. leguminosarum phaseoli,Burkholderiales optionally symbionts of Mimosa, Sinorhizobium meliloti,Sinorhizobium medicae, Sinorhizobium fredii, Sinorhizobium NGR234,Azorhizobium caulinodans, Bradyrhizobium japonicum, Bradyrhizobiumelkanii, Bradyrhizobium liaonginense, Frankia spp., and any combinationthereof, or by or by mycorrhizal fungi selected from the groupconsisting of Acaulosporaceae spp., Diversisporaceae spp., Gigasporaceaespp., Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagusspp., Sclerocystis spp., Septoglomus spp., Claroideoglomus spp.,Ambispora spp., Archaeospora spp., Geosiphon pyriformis, Paraglomusspp., other species in the division Glomeromycota, and any combinationthereof. In some embodiments, the modified polypeptide is localized to aplant cell plasma membrane. In some embodiments, the plant cell is aroot cell. In some embodiments, the root cell is a root epidermal cell.In some embodiments, the root cell is a root cortex cell. In someembodiments, the modified polypeptide is expressed in a developing plantroot system. In some embodiments, the first nucleic acid or secondnucleic acid sequence is operably linked to a promoter. In someembodiments, the promoter is a root specific promoter. In someembodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO:24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter, a derivative of the CaMV35Spromoter, the maize ubiquitin promoter, the trefoil promoter, a veinmosaic cassava virus promoter, or the Arabidopsis UBQ10 promoter. Insome embodiments, the plant is selected from the group of corn (e.g.,maize, Zea mays), rice (e.g., Oryza sativa, Oryza glaberrima, Zizaniaspp.), barley (e.g., Hordeum vulgare), wheat (e.g., common wheat, spelt,durum, Triticum aestivum, Triticum spelta, Triticum durum, Triticumspp.), Trema spp. (e.g., Trema cannabina, Trema cubense, Trema discolor,Trema domingensis, Trema integerrima, Trema lamarckiana, Tremamicrantha, Trema orientalis, Trema philippinensis, Trema strigilosa,Trema tomentosa), apple (e.g., Malus pumila), pear (e.g., Pyruscommunis, Pyrus×bretschneideri, Pyrus pyrifolia, Pyrus sinkiangensis,Pyrus pashia, Pyrus spp.), plum (e.g., prune, damson, bullaces, Prunusdomestica, Prunus salicina), apricot (e.g., Prunus armeniaca, Prunusbrigantina, Prunus mandshurica, Prunus mume, Prunus sibirica), peach(e.g., nectarine, Prunus persica), almond (e.g., Prunus dulcis, Prunusamygdalus), walnut (e.g., Persian walnut, English walnut, black walnut,Juglans regia, Juglans nigra, Juglans cinerea, Juglans californica),strawberry (e.g., Fragaria×ananassa, Fragaria chiloensis, Fragariavirginiana, Fragaria vesca), raspberry (e.g., European red raspberry,black raspberry, Rubus idaeus, Rubus occidentalis, Rubus strigosus),blackberry (e.g., evergreen blackberry, Himalayan blackberry, Rubusfruticosus, Rubus ursinus, Rubus laciniatus, Rubus argutus, Rubusarmeniacus, Rubus plicatus, Rubus ulmifolius, Rubus allegheniensis), redcurrant (e.g., Ribes rubrum, Ribes spicatum, Ribesbes alpinum, Ribesschlechtendalii, Ribes multiflorum, Ribes petraeum, Ribes triste), blackcurrant (e.g., Ribes nigrum), melon (e.g., watermelon, winter melon,casabas, cantaloupe, honeydew, muskmelon, Citrullus lanatus, Benincasahispida, Cucumis melo cantalupensis, Cucumis melo inodorus, Cucumis meloreticulatus), cucumber (e.g., slicing cucumbers, pickling cucumbers,English cucumber, Cucumis sativus), pumpkin (e.g., Cucurbita pepo,Cucurbita maxima), squash (e.g., gourd, Cucurbita argyrosperma,Cucurbita ficifolia, Cucurbita maxima, Cucurbita moschata), grape (e.g.,Vitis vinifera, Vitis amurensis, Vitis labrusca, Vitis mustangensis,Vitis riparia, Vitis rotundifolia), bean (e.g., Phaseolus vulgaris,Phaseolus lunatus, Vigna angularis, Vigna radiate, Vigna mungo,Phaseolus coccineus, Vigna umbellata, Vigna acontifolia, Phaseolusacutifolius, Vicia faba, Vicia faba equine, Phaseolus spp., Vigna spp.),soybean (e.g., soy, soya bean, Glycine max, Glycine soja), pea (e.g.,Pisum spp., Pisum sativum var. sativum, Pisum sativum var. arvense),chickpea (e.g., garbanzo, Bengal gram, Cicer arietinum), cowpea (e.g.,black-eyed pea, blackeye bean, Vigna unguiculata), pigeon pea (e.g.,Arhar/Toor, cajan pea, Congo bean, gandules, Caganus cajan), lentil(e.g., Lens culinaris), Bambara groundnut (e.g., earth pea, Vignasubterranea), lupin (e.g., Lupinus spp.), pulses (e.g., minor pulses,Lablab purpureaus, Canavalia ensiformis, Canavalia gladiate,Psophocarpus tetragonolobus, Mucuna pruriens var. utilis, Pachyrhizuserosus), Medicago spp. (e.g., Medicago sativa, Medicago truncatula,Medicago arborea), Lotus spp. (e.g., Lotus japonicus), forage legumes(e.g., Leucaena spp., Albizia spp., Cyamopsis spp., Sesbania spp.,Stylosanthes spp., Trifolium spp., Vicia spp.), indigo (e.g., Indigoferaspp., Indigofera tinctoria, Indigofera suffruticosa, Indigoferaarticulata, Indigofera oblongifolia, Indigofera aspalthoides, Indigoferasuffruticosa, Indigofera arrecta), legume trees (e.g., locust trees,Gleditsia spp., Robinia spp., Kentucky coffeetree, Gymnocladus dioicus,Acacia spp., Laburnum spp., Wisteria spp.), or hemp (e.g., cannabis,Cannabis sativa). In some embodiments, the plant part is a leaf, a stem,a root, a root primordia, a flower, a seed, a fruit, a kernel, a grain,a cell, or a portion thereof. In some embodiments, the plant part is afruit, a kernel, or a grain.

In some aspects, the present disclosure relates to a pollen grain or anovule of a genetically altered plant of any of the above embodimentsrelating to plants.

In some aspects, the present disclosure relates to a protoplast from agenetically altered plant of any of the above embodiments relating toplants.

In some aspects, the present disclosure relates to a tissue cultureproduced from protoplasts or cells from a genetically altered plant ofany of the above embodiments relating to plants, wherein the cells orprotoplasts are produced from a plant part selected from the groupconsisting of leaf, anther, pistil, stem, petiole, root, root primordia,root tip, fruit, seed, flower, cotyledon, hypocotyl, embryo, andmeristematic cell.

Methods of Producing and Cultivating Genetically Altered Plants

Certain aspects of the present disclosure relate to a method ofproducing the genetically altered plant of any one of the aboveembodiments relating to plants as described in the section “Geneticallyaltered plants and seeds”, comprising introducing a genetic alterationto the plant comprising the nucleic acid sequence. In some embodiments,the nucleic acid sequence, the first nucleic acid sequence, and/or thesecond nucleic acid sequence is operably linked to a promoter. In someembodiments, the promoter is a root specific promoter. In someembodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO:24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter (KAY et al. Science, 236, 4805,1987), a derivative of the CaMV35S promoter, the maize ubiquitinpromoter, the trefoil promoter, a vein mosaic cassava virus promoter, orthe Arabidopsis UBQ10 promoter. In some embodiments, the nucleic acidsequence, the first nucleic acid sequence, and/or the second nucleicacid sequence is inserted into the genome of the plant so that thenucleic acid sequence, the first nucleic acid sequence, and/or thesecond nucleic acid sequence is operably linked to an endogenouspromoter. In some embodiments, the endogenous promoter is a rootspecific promoter.

In some aspects, the present disclosure relates to a method of producinga genetically altered plant able to recognize LCOs, comprising the stepsof: introducing a genetic alteration to the plant comprising theprovision of an ability for LCOs produced by nitrogen-fixing bacteriaand/or mycorrhizal fungi to be recognized, thereby enabling the plant torecognize LCOs.

In some aspects, the present disclosure relates to a method of producinga genetically altered plant able to recognize LCOs, comprising the stepsof: introducing a genetic alteration to the plant comprising theprovision of an ability for LCOs produced by nitrogen-fixing bacteriaand/or mycorrhizal fungi to be recognized with high affinity, highselectivity, and/or high specificity, thereby enabling the plant torecognize LCOs with high affinity, high selectivity, and/or highspecificity.

In some aspects, the present disclosure relates to a method of producinga genetically altered plant able to recognize LCOs produced by aspecific nitrogen-fixing bacterial species and/or a specific mycorrhizalfungal species, comprising the steps of: introducing a geneticalteration to the plant comprising the provision of an ability for LCOsproduced by the specific nitrogen-fixing bacterial species and/or thespecific mycorrhizal fungal species to be recognized with alteredspecificity, thereby enabling the plant to recognize LCOs with alteredspecificity. In some embodiments, the genetic alteration allows thegenetically altered plant to recognize a different specificnitrogen-fixing bacterial species and/or specific mycorrhizal fungalspecies as compared to a plant without the genetic alteration. In someembodiments, the genetically altered plant is able to be grown indifferent agricultural conditions (e.g., different soils containingdifferent symbiotic microbial species, etc.). In some embodiments, thegenetic alteration allows the genetically altered plant to be grown indifferent agricultural conditions containing specific bacterial strainsproducing LCOs detected with high specificity, sensitivity, and/orselectivity by the genetically altered plant. In some embodiments, thebacterial strains are added as a seed coating or as a soil inoculum. Insome embodiments, the genetically altered plant is able to be grown withdifferent crop species (e.g., different crop rotations, etc.).

In some aspects, the present disclosure relates to a method ofcultivating a plant with the ability to recognize LCOs, comprising thesteps of: providing a seed with one or more genetic alterations thatprovide an ability for LCOs produced by nitrogen-fixing bacteria and/ormycorrhizal fungi to be recognized, wherein the seed produces a plantwith the ability to recognize LCOs produced by nitrogen-fixing bacteriaand/or mycorrhizal fungi; cultivating the plant under conditions wherethe ability to recognize LCOs produced by nitrogen-fixing bacteriaand/or mycorrhizal fungi results in increased growth, yield, and/orbiomass, as compared to a plant grown under the same conditions thatlacks the one or more genetic alterations. In some embodiments, theplant is cultivated in nutrient-poor soil.

In some aspects, the present disclosure relates to a method ofcultivating a plant with the ability to recognize LCOs with highaffinity, high selectivity, and/or high specificity, comprising thesteps of: providing a seed with one or more genetic alterations thatprovide an ability for LCOs produced by nitrogen-fixing bacteria and/ormycorrhizal fungi to be recognized with high affinity, high selectivity,and/or high specificity, wherein the seed produces a plant with theability to recognize LCOs produced by nitrogen-fixing bacteria and/ormycorrhizal fungi with high affinity, high selectivity, and/or highspecificity; cultivating the plant under conditions where the ability torecognize LCOs produced by nitrogen-fixing bacteria and/or mycorrhizalfungi with high affinity, high selectivity, and/or high specificityresults in increased growth, yield, and/or biomass, as compared to aplant grown under the same conditions that lacks the one or more geneticalterations. In some embodiments, the plant is cultivated innutrient-poor soil.

In some aspects, the present disclosure relates to a method ofcultivating a plant with the ability to recognize LCOs with alteredspecificity, comprising the steps of: providing a seed with one or moregenetic alterations that provide an ability for LCOs produced bynitrogen-fixing bacteria and/or mycorrhizal fungi to be recognized withaltered specificity, wherein the seed produces a plant with the abilityto recognize LCOs produced by nitrogen-fixing bacteria and/ormycorrhizal fungi with high specificity; cultivating the plant underconditions where the ability to recognize LCOs produced bynitrogen-fixing bacteria and/or mycorrhizal fungi with alteredspecificity results in increased growth, yield, and/or biomass, ascompared to a plant grown under the same conditions that lacks the oneor more genetic alterations. In some embodiments, the plant iscultivated in nutrient-poor soil. In some embodiments, the geneticalteration allows the genetically altered plant to recognize a differentspecific nitrogen-fixing bacterial species and/or specific mycorrhizalfungal species as compared to a plant without the genetic alteration. Insome embodiments, the genetically altered plant is able to be grown indifferent agricultural conditions (e.g., different soils containingdifferent symbiotic microbial species, etc.). In some embodiments, thegenetically altered plant is able to be grown with different cropspecies (e.g., different crop rotations, etc.).

In some aspects, the present disclosure relates to a method ofcultivating a plant with the ability to recognize LCOs, comprising thesteps of: providing a tissue culture or protoplast with one or moregenetic alterations that provide an ability for LCOs produced bynitrogen-fixing bacteria and/or mycorrhizal fungi to be recognized;regenerating the tissue culture or protoplast into a plantlet; growingthe plantlet into a plant, wherein the plant has the ability torecognize LCOs produced by nitrogen-fixing bacteria and/or mycorrhizalfungi; transplanting the plant into conditions where the ability torecognize LCOs produced by nitrogen-fixing bacteria and/or mycorrhizalfungi results in increased growth, yield, and/or biomass, as compared toa plant grown under the same conditions that lacks the one or moregenetic alterations. In some embodiments, the plant is cultivated innutrient-poor soil.

In some aspects, the present disclosure relates to a method ofcultivating a plant with the ability to recognize LCOs with highaffinity, high selectivity, and/or high specificity, comprising thesteps of: providing a tissue culture or protoplast with one or moregenetic alterations that provide an ability for LCOs produced bynitrogen-fixing bacteria and/or mycorrhizal fungi to be recognized withhigh affinity, high selectivity, and/or high specificity, regeneratingthe tissue culture or protoplast into a plantlet; growing the plantletinto a plant, wherein the plant has the ability to recognize LCOsproduced by produced by nitrogen-fixing bacteria and/or mycorrhizalfungi with high affinity, high selectivity, and/or high specificity;transplanting the plant into conditions where the ability to recognizeLCOs produced by nitrogen-fixing bacteria and/or mycorrhizal fungi withhigh affinity, high selectivity, and/or high specificity results inincreased growth, yield, and/or biomass, as compared to a plant grownunder the same conditions that lacks the one or more geneticalterations. In some embodiments, the plant is cultivated innutrient-poor soil.

In some aspects, the present disclosure relates to a method ofcultivating a plant with the ability to recognize LCOs with alteredspecificity, comprising the steps of: providing a tissue culture orprotoplast with one or more genetic alterations that provide an abilityfor LCOs produced by nitrogen-fixing bacteria and/or mycorrhizal fungito be recognized with altered specificity, regenerating the tissueculture or protoplast into a plantlet; growing the plantlet into aplant, wherein the plant has the ability to recognize LCOs produced byproduced by nitrogen-fixing bacteria and/or mycorrhizal fungi with highspecificity; transplanting the plant into conditions where the abilityto recognize LCOs produced by nitrogen-fixing bacteria and/ormycorrhizal fungi with altered specificity results in increased growth,yield, and/or biomass, as compared to a plant grown under the sameconditions that lacks the one or more genetic alterations. In someembodiments, the plant is cultivated in nutrient-poor soil. In someembodiments, the genetic alteration allows the genetically altered plantto recognize a different specific nitrogen-fixing bacterial speciesand/or specific mycorrhizal fungal species as compared to a plantwithout the genetic alteration. In some embodiments, the geneticallyaltered plant is able to be grown in different agricultural conditions(e.g., different soils containing different symbiotic microbial species,etc.). In some embodiments, the genetically altered plant is able to begrown with different crop species (e.g., different crop rotations,etc.).

In some embodiments of any of the above methods, the ability torecognize LCOs is conferred by a nucleic acid sequence encoding amodified plant LysM receptor of any one of the embodiments described inthe section “Modified plant LysM receptors”. In some embodiments, themodified plant LysM receptor has higher selectivity and/or affinity forLCOs than the unmodified plant LysM receptor and the expression of themodified plant LysM receptor allows the plant or part thereof torecognize LCOs with high selectivity and/or affinity. In someembodiments, the LCOs are produced by nitrogen-fixing bacteria or bymycorrhizal fungi. In some embodiments, the LCOs are produced bynitrogen-fixing bacteria selected from the group consisting ofMesorhizobium loti, Mesorhizobium huakuii, Mesorhizobium mediterraneum,Mesorhizobium ciceri, Mesorhizobium spp., Rhizobium mongolense,Rhizobium tropici, Rhizobium etli phaseoli, Rhizobium giardinii,Rhizobium leguminosarum optionally R. leguminosarum trifolii, R.leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., and any combination thereof,or by or by mycorrhizal fungi selected from the group consisting ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, and any combination thereof. Insome embodiments, the modified polypeptide is localized to a plant cellplasma membrane. In some embodiments, the plant cell is a root cell. Insome embodiments, the root cell is a root epidermal cell. In someembodiments, the root cell is a root cortex cell. In some embodiments,the modified polypeptide is expressed in a developing plant root system.In some embodiments, the nucleic acid sequence is operably linked to apromoter. In some embodiments, the promoter is a root specific promoter.In some embodiments, the promoter is selected from the group of aNFR1/LYK3/CERK6 or NFR5/NFP promoter, the Lotus NFR5 promoter (SEQ IDNO:24) and the Lotus NFR1 promoters (SEQ ID NO:25) the maizeallothioneine promoter, the chitinase promoter, the maize ZRP2 promoter,the tomato LeExt1 promoter, the glutamine synthetase soybean rootpromoter, the RCC3 promoter, the rice antiquitine promoter, the LRRreceptor kinase promoter, or the Arabidopsis pCO2 promoter. In someembodiments, the promoter is a constitutive promoter optionally selectedfrom the group of the CaMV35S promoter, a derivative of the CaMV35Spromoter, the maize ubiquitin promoter, the trefoil promoter, a veinmosaic cassava virus promoter, or the Arabidopsis UBQ10 promoter. Insome embodiments, the plant is selected from the group of corn (e.g.,maize, Zea mays), rice (e.g., Oryza sativa, Oryza glaberrima, Zizaniaspp.), barley (e.g., Hordeum vulgare), wheat (e.g., common wheat, spelt,durum, Triticum aestivum, Triticum spelta, Triticum durum, Triticumspp.), Trema spp. (e.g., Trema cannabina, Trema cubense, Trema discolor,Trema domingensis, Trema integerrima, Trema lamarckiana, Tremamicrantha, Trema orientalis, Trema philippinensis, Trema strigilosa,Trema tomentosa), apple (e.g., Malus pumila), pear (e.g., Pyruscommunis, Pyrus xbretschneideri, Pyrus pyrifolia, Pyrus sinkiangensis,Pyrus pashia, Pyrus spp.), plum (e.g., prune, damson, bullaces, Prunusdomestica, Prunus salicina), apricot (e.g., Prunus armeniaca, Prunusbrigantina, Prunus mandshurica, Prunus mume, Prunus sibirica), peach(e.g., nectarine, Prunus persica), almond (e.g., Prunus dulcis, Prunusamygdalus), walnut (e.g., Persian walnut, English walnut, black walnut,Juglans regia, Juglans nigra, Juglans cinerea, Juglans californica),strawberry (e.g., Fragaria×ananassa, Fragaria chiloensis, Fragariavirginiana, Fragaria vesca), raspberry (e.g., European red raspberry,black raspberry, Rubus idaeus, Rubus occidentalis, Rubus strigosus),blackberry (e.g., evergreen blackberry, Himalayan blackberry, Rubusfruticosus, Rubus ursinus, Rubus laciniatus, Rubus argutus, Rubusarmeniacus, Rubus plicatus, Rubus ulmifolius, Rubus allegheniensis), redcurrant (e.g., Ribes rubrum, Ribes spicatum, Ribesbes alpinum, Ribesschlechtendalii, Ribes multiflorum, Ribes petraeum, Ribes triste), blackcurrant (e.g., Ribes nigrum), melon (e.g., watermelon, winter melon,casabas, cantaloupe, honeydew, muskmelon, Citrullus lanatus, Benincasahispida, Cucumis melo cantalupensis, Cucumis melo inodorus, Cucumis meloreticulatus), cucumber (e.g., slicing cucumbers, pickling cucumbers,English cucumber, Cucumis sativus), pumpkin (e.g., Cucurbita pepo,Cucurbita maxima), squash (e.g., gourd, Cucurbita argyrosperma,Cucurbita ficifolia, Cucurbita maxima, Cucurbita moschata), grape (e.g.,Vitis vinifera, Vitis amurensis, Vitis labrusca, Vitis mustangensis,Vitis riparia, Vitis rotundifolia), bean (e.g., Phaseolus vulgaris,Phaseolus lunatus, Vigna angularis, Vigna radiate, Vigna mungo,Phaseolus coccineus, Vigna umbellate, Vigna acontifolia, Phaseolusacutifolius, Vicia faba, Vicia faba equine, Phaseolus spp., Vigna spp.),soybean (e.g., soy, soya bean, Glycine max, Glycine soja), pea (e.g.,Pisum spp., Pisum sativum var. sativum, Pisum sativum var. arvense),chickpea (e.g., garbanzo, Bengal gram, Cicer arietinum), cowpea (e.g.,black-eyed pea, blackeye bean, Vigna unguiculata), pigeon pea (e.g.,Arhar/Toor, cajan pea, Congo bean, gandules, Caganus cajan), lentil(e.g., Lens culinaris), Bambara groundnut (e.g., earth pea, Vignasubterranea), lupin (e.g., Lupinus spp.), pulses (e.g., minor pulses,Lablab purpureaus, Canavalia ensiformis, Canavalia gladiate,Psophocarpus tetragonolobus, Mucuna pruriens var. utilis, Pachyrhizuserosus), Medicago spp. (e.g., Medicago sativa, Medicago truncatula,Medicago arborea), Lotus spp. (e.g., Lotus japonicus), forage legumes(e.g., Leucaena spp., Albizia spp., Cyamopsis spp., Sesbania spp.,Stylosanthes spp., Trifolium spp., Vicia spp.), indigo (e.g., Indigoferaspp., Indigofera tinctoria, Indigofera suffruticosa, Indigoferaarticulata, Indigofera oblongifolia, Indigofera aspalthoides, Indigoferasuffruticosa, Indigofera arrecta), legume trees (e.g., locust trees,Gleditsia spp., Robinia spp., Kentucky coffeetree, Gymnocladus dioicus,Acacia spp., Laburnum spp., Wisteria spp.), or hemp (e.g., cannabis,Cannabis sativa).

Molecular Biological Methods to Produce Genetically Altered Plants andPlant Cells

One embodiment of the present invention provides a genetically alteredplant or plant cell comprising one or more modified endogenous plantgenes. For example, the present disclosure provides plants withgenetically altered LysM receptors modified to include a hydrophobicpatch or alter the hydrophobic patch in the LysM2 domain and plants withgenetically altered LysM receptors modified by replacing regions in theLysM1 domain with corresponding donor LysM1 domain regions. Plants withthese modified receptors can have increased affinity, selectivity,and/or specificity for LCOs.

Certain aspects of the present disclosure relate to methods forselection of a target plant LysM receptor for modifying the target plantLysM receptor to have a desired receptor characteristic, wherein themethod includes the steps of: a) providing a structural model, amolecular model, a surface characteristics model, and/or anelectrostatic potential model of a donor plant LysM receptor having thedesired receptor characteristic and two or more potential target plantLysM receptors; b) comparing each of the two or more potential targetplant LysM receptors with the structural model, the molecular model, thesurface characteristics model, and/or the electrostatic potential modelof the donor plant LysM receptor, and/or comparing each of the two ormore potential target plant LysM receptors with the donor plant LysMreceptor using structural overlay; and c) selecting the potential targetplant LysM receptor with a suitable match for the donor plant LysMreceptor to be the target plant LysM receptor. In some embodiments, thecriteria for determining that the potential target plant LysM receptoris a suitable match for the donor plant LysM receptor in step (c) areselected from the group of goodness of fit to template structure;similarity; phylogenetic relation; surface potential; coverage totemplate structure; GMQE, QMEAN, and Local Quality estimates fromSWISS-Model; or any combination thereof. In some embodiments, thestructural model of a donor plant LysM receptor is a protein crystalstructure, a molecular model, a cryo-EM structure, and a NMR structure.In some embodiments, the donor plant LysM receptor model is of an entireectodomain and the two or more potential target plant LysM receptormodels are of entire ectodomains. In some embodiments, the donor plantLysM receptor model is of a LysM1 domain, a LysM2 domain, a LysM3domain, or any combination thereof, and the two or more potential targetplant LysM receptor models are of LysM1 domains, LysM2 domains, LysM3domains, or any combination thereof.

In some embodiments, the donor plant LysM receptor is Medicago NFP,Medicago LYK3, Lotus NFR1, Lotus NFR5, Lotus LYS11, or ArabidopsisCERK1. In some embodiments, the two or more target plant LysM receptorsare additionally compared to Lotus CERK6. In some embodiments, the twoor more potential target plant LysM receptor polypeptides are all fromthe same plant species or plant variety. In some embodiments, thedesired receptor characteristic is affinity, selectivity, and/orspecificity for an oligosaccharide or class of oligosaccharides. In someembodiments, the desired receptor characteristic is binding kinetics foran oligosaccharide or class of oligosaccharides, wherein the bindingkinetics include off-rate and on-rate. In some embodiments, the class ofoligosaccharides is selected from the group of LCOs, COs, beta-glucans,cyclic-beta-glucans, exopolysaccharides, or optionally LPS. In someembodiments, the class of oligosaccharides is LCOs or COs. In someembodiments, the class of oligosaccharides is LCOs, optionally producedby a produced by a nitrogen-fixing bacteria optionally selected from thegroup of Mesorhizobium loti, Mesorhizobium huakuii, Mesorhizobiummediterraneum, Mesorhizobium ciceri, Mesorhizobium spp., Rhizobiummongolense, Rhizobium tropici, Rhizobium etli phaseoli, Rhizobiumgiardinii, Rhizobium leguminosarum optionally R. leguminosarum trifolii,R. leguminosarum viciae, and R. leguminosarum phaseoli, Burkholderialesoptionally symbionts of Mimosa, Sinorhizobium meliloti, Sinorhizobiummedicae, Sinorhizobium fredii, Sinorhizobium NGR234, Azorhizobiumcaulinodans, Bradyrhizobium japonicum, Bradyrhizobium elkanii,Bradyrhizobium liaonginense, Frankia spp., or any combination thereof;or optionally produced by a mycorrhizal fungi optionally selected fromthe group of Acaulosporaceae spp., Diversisporaceae spp., Gigasporaceaespp., Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagusspp., Sclerocystis spp., Septoglomus spp., Claroideoglomus spp.,Ambispora spp., Archaeospora spp., Geosiphon pyriformis, Paraglomusspp., other species in the division Glomeromycota, or any combinationthereof. In some embodiments, the LCOs are M. loti LCO, S. melilotiLCO-IV, or S. meliloti LCO-V.

In some embodiments, the method further includes step d) identifying oneor more amino acid residues for modification in the target LysM receptorby comparing amino acid residues of a first oligosaccharide bindingfeature in the donor plant LysM receptor with the corresponding aminoacid residues in the target plant LysM receptor, and optionallyidentifying one or more amino acid residues for modification in thetarget LysM receptor by comparing amino acid residues of a secondoligosaccharide binding feature in the donor plant LysM receptor withthe corresponding amino acid residues in the target plant LysM receptor.In some embodiments, the method further includes step e) generating amodified plant LysM receptor wherein the one or more amino acid residuesin the first oligosaccharide binding feature of the target plant LysMreceptor have been substituted with corresponding amino acid residuesfrom the donor LysM; generating a modified plant LysM receptor whereinthe one or more amino acid residues in the second oligosaccharidebinding feature of the target plant LysM receptor have been substitutedwith corresponding amino acid residues from the donor LysM; orgenerating a modified plant LysM receptor wherein the one or more aminoacid residues in the first oligosaccharide binding feature of the targetplant LysM receptor have been substituted with corresponding amino acidresidues from the donor LysM and the one or more amino acid residues inthe second oligosaccharide binding feature of the target plant LysMreceptor have been substituted with corresponding amino acid residuesfrom the donor LysM. In some embodiments, the first oligosaccharidebinding feature is a hydrophobic patch on the surface of the LysM2domain. In some embodiments, the second oligosaccharide binding featureis a part of the LysM1 domain of the donor plant LysM receptor. In someembodiments, the modified LysM receptor is an endogenous LysM receptormodified using one or more gene editing components. In some embodiments,the one or more gene editing components target a nuclear genome sequenceoperably linked to a nucleic acid encoding an endogenous LysM receptor(e.g., a soybean LysM receptor). A further embodiment of this aspectincludes the one or more gene editing components being selected from thegroup of a ribonucleoprotein complex that targets the nuclear genomesequence; a vector comprising a TALEN protein encoding sequence, whereinthe TALEN protein targets the nuclear genome sequence; a vectorcomprising a ZFN protein encoding sequence, wherein the ZFN proteintargets the nuclear genome sequence; an oligonucleotide donor (ODN),wherein the ODN targets the nuclear genome sequence; or a vectorcomprising a CRISPR/Cas enzyme encoding sequence and a targetingsequence, wherein the targeting sequence targets the nuclear genomesequence.

Additional aspects of the present disclosure relate to a modified plantLysM receptor produced using any one of the above methods, wherein themodified plant LysM receptor includes a LysM2 domain modified tocomprise a hydrophobic patch on the surface of the LysM2 domain. Furtheraspects of the present disclosure relate to a modified plant LysMreceptor produced using any one of the above methods, wherein themodified plant LysM receptor includes a first LysM1 domain modified toreplace at least part of the first LysM1 domain with at least part of asecond LysM1 domain. The modified plant LysM receptors of the presentdisclosure may be used to produce the genetically altered plant of anyone of the above embodiments relating to plants as described in thesection “Genetically altered plants and seeds”

Transformation and generation of genetically altered monocotyledonousand dicotyledonous plant cells is well known in the art. See, e.g.,Weising, et al., Ann. Rev. Genet. 22:421-477 (1988); U.S. Pat. No.5,679,558; Agrobacterium Protocols, ed: Gartland, Humana Press Inc.(1995); and Wang, et al. Acta Hort. 461:401-408 (1998). The choice ofmethod varies with the type of plant to be transformed, the particularapplication and/or the desired result. The appropriate transformationtechnique is readily chosen by the skilled practitioner.

Any methodology known in the art to delete, insert or otherwise modifythe cellular DNA (e.g., genomic DNA and organelle DNA) can be used inpracticing the inventions disclosed herein. For example, a disarmed Tiplasmid, containing a genetic construct for deletion or insertion of atarget gene, in Agrobacterium tumefaciens can be used to transform aplant cell, and thereafter, a transformed plant can be regenerated fromthe transformed plant cell using procedures described in the art, forexample, in EP 0116718, EP 0270822, PCT publication WO 84/02913 andpublished European Patent application (“EP”) 0242246. Ti-plasmid vectorseach contain the gene between the border sequences, or at least locatedto the left of the right border sequence, of the T-DNA of theTi-plasmid. Of course, other types of vectors can be used to transformthe plant cell, using procedures such as direct gene transfer (asdescribed, for example in EP 0233247), pollen mediated transformation(as described, for example in EP 0270356, PCT publication WO 85/01856,and U.S. Pat. No. 4,684,611), plant RNA virus-mediated transformation(as described, for example in EP 0 067 553 and U.S. Pat. No. 4,407,956),liposome-mediated transformation (as described, for example in U.S. Pat.No. 4,536,475), and other methods such as the methods for transformingcertain lines of corn (e.g., U.S. Pat. No. 6,140,553; Fromm et al.,Bio/Technology (1990) 8, 833 839); Gordon-Kamm et al., The Plant Cell,(1990) 2, 603 618) and rice (Shimamoto et al., Nature, (1989) 338, 274276; Datta et al., Bio/Technology, (1990) 8, 736 740) and the method fortransforming monocots generally (PCT publication WO 92/09696). Forcotton transformation, the method described in PCT patent publication WO00/71733 can be used. For soybean transformation, reference is made tomethods known in the art, e.g., Hinchee et al. (Bio/Technology, (1988)6, 915) and Christou et al. (Trends Biotech, (1990) 8, 145) or themethod of WO 00/42207.

Genetically altered plants of the present invention can be used in aconventional plant breeding scheme to produce more genetically alteredplants with the same characteristics, or to introduce the geneticalteration(s) in other varieties of the same or related plant species.Seeds, which are obtained from the altered plants, preferably containthe genetic alteration(s) as a stable insert in chromosomal or organelleDNA or as modifications to an endogenous gene or promoter. Plantscomprising the genetic alteration(s) in accordance with the inventioninclude plants comprising, or derived from, root stocks of plantscomprising the genetic alteration(s) of the invention, e.g., fruit treesor ornamental plants. Hence, any non-transgenic grafted plant partsinserted on a transformed plant or plant part are included in theinvention.

Introduced genetic elements, whether in an expression vector orexpression cassette, which result in the expression of an introducedgene will typically utilize a plant-expressible promoter. A‘plant-expressible promoter’ as used herein refers to a promoter thatensures expression of the genetic alteration(s) of the invention in aplant cell. Examples of promoters directing constitutive expression inplants are known in the art and include: the strong constitutive 35Spromoters (the “35S promoters”) of the cauliflower mosaic virus (CaMV),e.g., of isolates CM 1841 (Gardner et al., Nucleic Acids Res, (1981) 9,2871 2887), CabbB S (Franck et al., Cell (1980) 21, 285 294) and CabbB JI (Hull and Howell, Virology, (1987) 86, 482 493); promoters from theubiquitin family (e.g., the maize ubiquitin promoter of Christensen etal., Plant Mol Biol, (1992) 18, 675-689), the gos2 promoter (de Pater etal., The Plant J (1992) 2, 834-844), the emu promoter (Last et al.,Theor Appl Genet, (1990) 81, 581-588), actin promoters such as thepromoter described by An et al. (The Plant J, (1996) 10, 107), the riceactin promoter described by Zhang et al. (The Plant Cell, (1991) 3,1155-1165); promoters of the Cassava vein mosaic virus (WO 97/48819,Verdaguer et al. (Plant Mol Biol, (1998) 37, 1055-1067), the pPLEXseries of promoters from Subterranean Clover Stunt Virus (WO 96/06932,particularly the S4 or S7 promoter), an alcohol dehydrogenase promoter,e.g., pAdh1S (GenBank accession numbers X04049, X00581), and the TR1′promoter and the TR2′ promoter (the “TR1′ promoter” and “TR2′ promoter”,respectively) which drive the expression of the 1′ and 2′ genes,respectively, of the T DNA (Velten et al., EMBO J, (1984) 3, 2723 2730).

Alternatively, a plant-expressible promoter can be a tissue-specificpromoter, i.e., a promoter directing a higher level of expression insome cells or tissues of the plant, e.g., in root epidermal cells orroot cortex cells. Examples of constitutive promoters that are oftenused in plant cells are the cauliflower mosaic (CaMV) 35S promoter (KAYet al. Science, 236, 4805, 1987), and various derivatives of thepromoter, virus promoter vein mosaic cassava (International ApplicationWO 97/48819), the maize ubiquitin promoter (CHRISTENSEN & QUAIL,Transgenic Res, 5, 213-8, 1996), trefoil (Ljubql, MAEKAWA et al. MolPlant Microbe Interact. 21, 375-82, 2008) and Arabidopsis (UBQ10, Norriset al. Plant Mol. Biol. 21, 895-906, 1993).

In preferred embodiments, root specific promoters will be used.Non-limiting examples include the promoter of the maize allothioneine(DE FRAMOND et al, FEBS 290, 103-106, 1991 Application EP 452269), thechitinase promoter (SAMAC et al. Plant Physiol 93, 907-914, 1990), theglutamine synthetase soybean root promoter (HIREL et al. Plant Mol.Biol. 20, 207-218, 1992), the RCC3 promoter (PCT Application WO2009/016104), the rice antiquitine promoter (PCT Application WO2007/076115), the LRR receptor kinase promoter (PCT application WO02/46439), the maize ZRP2 promoter (U.S. Pat. No. 5,633,363), the tomatoLeExt1 promoter (Bucher et al. Plant Physiol. 128, 911-923, 2002), andthe Arabidopsis pCO2 promoter (HEIDSTRA et al, Genes Dev. 18, 1964-1969,2004). These plant promoters can be combined with enhancer elements,they can be combined with minimal promoter elements, or can compriserepeated elements to ensure the expression profile desired.

In some embodiments, genetic elements to increase expression in plantcells can be utilized. For example, an intron at the 5′ end or 3′ end ofan introduced gene, or in the coding sequence of the introduced gene,e.g., the hsp70 intron. Other such genetic elements can include, but arenot limited to, promoter enhancer elements, duplicated or triplicatedpromoter regions, 5′ leader sequences different from another transgeneor different from an endogenous (plant host) gene leader sequence, 3′trailer sequences different from another transgene used in the sameplant or different from an endogenous (plant host) trailer sequence.

An introduced gene of the present invention can be inserted in host cellDNA so that the inserted gene part is upstream (i.e., 5′) of suitable 3′end transcription regulation signals (e.g., transcript formation andpolyadenylation signals). This is preferably accomplished by insertingthe gene in the plant cell genome (nuclear or chloroplast). Preferredpolyadenylation and transcript formation signals include those of thenopaline synthase gene (Depicker et al., J. Molec Appl Gen, (1982) 1,561-573), the octopine synthase gene (Gielen et al., EMBO J, (1984)3:835 845), the SCSV or the Malic enzyme terminators (Schunmann et al.,Plant Funct Biol, (2003) 30:453-460), and the T DNA gene 7 (Velten andSchell, Nucleic Acids Res, (1985) 13, 6981 6998), which act as 3′untranslated DNA sequences in transformed plant cells. In someembodiments, one or more of the introduced genes are stably integratedinto the nuclear genome. Stable integration is present when the nucleicacid sequence remains integrated into the nuclear genome and continuesto be expressed (e.g., detectable mRNA transcript or protein isproduced) throughout subsequent plant generations. Stable integrationinto and/or editing of the nuclear genome can be accomplished by anyknown method in the art (e.g., microparticle bombardment,Agrobacterium-mediated transformation, CRISPR/Cas9, electroporation ofprotoplasts, microinjection, etc.).

The term recombinant or modified nucleic acids refers to polynucleotideswhich are made by the combination of two otherwise separated segments ofsequence accomplished by the artificial manipulation of isolatedsegments of polynucleotides by genetic engineering techniques or bychemical synthesis. In so doing one may join together polynucleotidesegments of desired functions to generate a desired combination offunctions.

As used herein, the terms “overexpression” and “upregulation” refer toincreased expression (e.g., of mRNA, polypeptides, etc.) relative toexpression in a wild type organism (e.g., plant) as a result of geneticmodification. In some embodiments, the increase in expression is aslight increase of about 10% more than expression in wild type. In someembodiments, the increase in expression is an increase of 50% or more(e.g., 60%, 70%, 80%, 100%, etc.) relative to expression in wild type.In some embodiments, an endogenous gene is overexpressed. In someembodiments, an exogenous gene is overexpressed by virtue of beingexpressed. Overexpression of a gene in plants can be achieved throughany known method in the art, including but not limited to, the use ofconstitutive promoters, inducible promoters, high expression promoters(e.g., PsaD promoter), enhancers, transcriptional and/or translationalregulatory sequences, codon optimization, modified transcriptionfactors, and/or mutant or modified genes that control expression of thegene to be overexpressed.

Where a recombinant nucleic acid is intended for expression, cloning, orreplication of a particular sequence, DNA constructs prepared forintroduction into a host cell will typically comprise a replicationsystem (e.g. vector) recognized by the host, including the intended DNAfragment encoding a desired polypeptide, and can also includetranscription and translational initiation regulatory sequences operablylinked to the polypeptide-encoding segment. Additionally, suchconstructs can include cellular localization signals (e.g., plasmamembrane localization signals). In preferred embodiments, such DNAconstructs are introduced into a host cell's genomic DNA, chloroplastDNA or mitochondrial DNA.

In some embodiments, a non-integrated expression system can be used toinduce expression of one or more introduced genes. Expression systems(expression vectors) can include, for example, an origin of replicationor autonomously replicating sequence (ARS) and expression controlsequences, a promoter, an enhancer and necessary processing informationsites, such as ribosome-binding sites, RNA splice sites, polyadenylationsites, transcriptional terminator sequences, and mRNA stabilizingsequences. Signal peptides can also be included where appropriate fromsecreted polypeptides of the same or related species, which allow theprotein to cross and/or lodge in cell membranes, cell wall, or besecreted from the cell.

Selectable markers useful in practicing the methodologies of theinvention disclosed herein can be positive selectable markers.Typically, positive selection refers to the case in which a geneticallyaltered cell can survive in the presence of a toxic substance only ifthe recombinant polynucleotide of interest is present within the cell.Negative selectable markers and screenable markers are also well knownin the art and are contemplated by the present invention. One of skillin the art will recognize that any relevant markers available can beutilized in practicing the inventions disclosed herein.

Screening and molecular analysis of recombinant strains of the presentinvention can be performed utilizing nucleic acid hybridizationtechniques. Hybridization procedures are useful for identifyingpolynucleotides, such as those modified using the techniques describedherein, with sufficient homology to the subject regulatory sequences tobe useful as taught herein. The particular hybridization techniques arenot essential to the subject invention. As improvements are made inhybridization techniques, they can be readily applied by one of skill inthe art. Hybridization probes can be labeled with any appropriate labelknown to those of skill in the art. Hybridization conditions and washingconditions, for example temperature and salt concentration, can bealtered to change the stringency of the detection threshold. See, e.g.,Sambrook et al. (1989) vide infra or Ausubel et al. (1995) CurrentProtocols in Molecular Biology, John Wiley & Sons, NY, N.Y., for furtherguidance on hybridization conditions.

Additionally, screening and molecular analysis of genetically alteredstrains, as well as creation of desired isolated nucleic acids can beperformed using Polymerase Chain Reaction (PCR). PCR is a repetitive,enzymatic, primed synthesis of a nucleic acid sequence. This procedureis well known and commonly used by those skilled in this art (seeMullis, U.S. Pat. Nos. 4,683,195, 4,683,202, and 4,800,159; Saiki et al.(1985) Science 230:1350-1354). PCR is based on the enzymaticamplification of a DNA fragment of interest that is flanked by twooligonucleotide primers that hybridize to opposite strands of the targetsequence. The primers are oriented with the 3′ ends pointing towardseach other. Repeated cycles of heat denaturation of the template,annealing of the primers to their complementary sequences, and extensionof the annealed primers with a DNA polymerase result in theamplification of the segment defined by the 5′ ends of the PCR primers.Because the extension product of each primer can serve as a template forthe other primer, each cycle essentially doubles the amount of DNAtemplate produced in the previous cycle. This results in the exponentialaccumulation of the specific target fragment, up to several million-foldin a few hours. By using a thermostable DNA polymerase such as the Taqpolymerase, which is isolated from the thermophilic bacterium Thermusaquaticus, the amplification process can be completely automated. Otherenzymes which can be used are known to those skilled in the art.

Nucleic acids and proteins of the present invention can also encompasshomologues of the specifically disclosed sequences. Homology (e.g.,sequence identity) can be 50%-100%. In some instances, such homology isgreater than 80%, greater than 85%, greater than 90%, or greater than95%. The degree of homology or identity needed for any intended use ofthe sequence(s) is readily identified by one of skill in the art. Asused herein percent sequence identity of two nucleic acids is determinedusing an algorithm known in the art, such as that disclosed by Karlinand Altschul (1990) Proc. Natl. Acad. Sci. USA 87:2264-2268, modified asin Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877.Such an algorithm is incorporated into the NBLAST and XBLAST programs ofAltschul et al. (1990) J. Mol. Biol. 215:402-410. BLAST nucleotidesearches are performed with the NBLAST program, score=100,wordlength=12, to obtain nucleotide sequences with the desired percentsequence identity. To obtain gapped alignments for comparison purposes,Gapped BLAST is used as described in Altschul et al. (1997) Nucl. Acids.Res. 25:3389-3402. When utilizing BLAST and Gapped BLAST programs, thedefault parameters of the respective programs (NBLAST and XBLAST) areused. See www.ncbi.nih.gov.

Preferred host cells are plant cells. Recombinant host cells, in thepresent context, are those which have been genetically modified tocontain an isolated nucleic molecule, contain one or more deleted orotherwise non-functional genes normally present and functional in thehost cell, or contain one or more genes to produce at least onerecombinant protein. The nucleic acid(s) encoding the protein(s) of thepresent invention can be introduced by any means known to the art whichis appropriate for the particular type of cell, including withoutlimitation, transformation, lipofection, electroporation or any othermethodology known by those skilled in the art.

Having generally described this invention, the same will be betterunderstood by reference to certain specific examples, which are includedherein to further illustrate the invention and are not intended to limitthe scope of the invention as defined by the claims.

EXAMPLES

The present disclosure is described in further detail in the followingexamples which are not in any way intended to limit the scope of thedisclosure as claimed. The attached figures are meant to be consideredas integral parts of the specification and description of thedisclosure. The following examples are offered to illustrate, but not tolimit the claimed disclosure.

Example 1: Structural Characterization of Medicago NFP Ectodomain

The following example describes the structural characterization of theMedicago NFP protein ectodomain.

Materials and Methods

Expression and purification of Medicago NFP ectodomain: The Medicagotruncatula NFP ectodomain (residues 28-246) was codon-optimized forinsect cell expression (Genscript, Piscataway, USA) and cloned into thepOET4 baculovirus transfer vector (Oxford Expression Technologies). Thenative NFP signal peptide (residues 1-27, predicted by SignalP 4.1) wasreplaced with the AcMNPV gp67 signal peptide to facilitate secretion anda hexa-histidine tag was added to the C-terminus. Recombinantbaculoviruses were produced in Sf9 cells (Spodoptera frugiperda) usingthe FlashBac Gold kit (Oxford Expression technologies) according to themanufacturer's instructions with Lipofectin (ThermoFisher Scientific) asa transfection reagent. Protein expression was performed as follows.Suspension-cultured Sf9 cells were maintained with shaking at 299 K inserum-free MAX-XP (BD-Biosciences, discontinued) or HyClone SFX (GEHealthcare) medium supplemented with 1% Pen-Strep (10000 U/ml, Lifetechnologies) and 1% CD lipid concentrate (Gibco). Protein expressionwas induced by adding recombinant passage 3 virus once the Sf9 cellsreached a cell density of 1.0*10{circumflex over ( )}6 cells/ml. After5-7 days of expression, medium supernatant containing NFP ectodomainswas harvested by centrifugation. This was followed by an overnightdialysis step against 50 mM Tris-HCl pH 8, 200 mM NaCl at 277 K. The NFPectodomain was enriched by two subsequent steps of Ni-IMAC purification(HisTrap excel/HisTrap HP, both GE Healthcare). For crystallography,N-glycans were removed using the endoglycosidase PNGase F (1:15 (w/w),room temperature, overnight). As a final purification step, NFPectodomains was purified by SEC on a Superdex 200 10/300 or HiLoadSuperdex 200 16/600 (both GE Healthcare) in phosphate buffered saline atpH 7.2 supplemented to a total of 500 mM NaCl (for binding assays) or 50mM Tris-HCl, 200 mM NaCl (for crystallography). NFP ectodomain elutes asa single, homogeneous peak corresponding to a monomer.

Crystallization and structure determination: Crystals of deglycosylatedNFP ectodomain were obtained using a vapour diffusion setup at 3-5 mg/mlin 0.2 M Na-acetate, 0.1 M Na-cacodylate pH 6.5, and 30% (w/v) PEG-8000.Crystals were cryoprotected in their crystallization condition bysupplementing with 5% (w/v) PEG-400 before being snap-frozen in liquidnitrogen. Complete diffraction data to 2.85 Å resolution was obtained atthe MaxLab 1911-3 beamline. The phase problem was solved by molecularreplacement using Phaser from the PHENIX suite with a homology modelbased on the AtCERK1 ectodomain structure (PDB coordinates 4EBZ) as asearch model. Model building and refinement was done using COOT and thePHENIX suite, respectively. The output pdb filled structural model wasgenerated and its electrostatic surface potential was calculated usingthe PDB2PQR and APBS webservers (PMID: 21425296). The results werevisualized in PyMol using APBS tools 2.1 (DeLano, W. L. (2002). PyMOL.DeLano Scientific, San Carlos, Calif., 700.).

Results

The structure of Medicago NFP was determined by molecular replacementusing a homology model based on the inner low B-factor scaffold ofAtCERK1. The complete structure of NFP (residues 33-233) was built thisway, including four N-glycosylations that were clearly resolved in the2.8 Å electron density map. NFP forms a compact structure where threeclassical βααβ LysM domains are tightly interconnected and stabilized by3 conserved disulfide bridges (C3-C104, C47-C166 and C102-C164) (FIG.1). The disulfide connectivity pattern and the overall scaffoldarrangement is shared with other LysM-RLK proteins involved in chitindefense signaling, supporting a common evolutionary origin of theseclass of receptors.

Example 2: Identification of Important Residues forLipo-Chitooligosaccharide (LCO) Perception

The following example describes the use of a structurally-guidedapproach to identify important residues in NFP for LCO perception. Afteridentifying important residues, NFP point mutations were created, andtested using ligand-binding assays.

Materials and Methods

Structurally-guided residue identification: The NFP ectodomain wasstructurally aligned to ligand-bound CERK1. Then, the electrostaticsurface potential was mapped to the previously-developed structure ofthe NFP ectodomain. The predicted ligand-binding location andelectrostatic surface potential are depicted in FIG. 2A.

Creation of NFP point mutations: The NFP leucine residues L147 and L154were replaced with aspartate residues. Aspartate is similar in size toleucine, but negatively charged where leucine is hydrophobic. Pointmutants of NFP were engineered using site-directed mutagenesis. Inparticular, a double-mutated NFP was engineered where the leucineresidues L147 and L154 were replaced with aspartate residues to createthe mutant NFP L147D L154D. Point mutated versions of NFP were expressedand purified as described in Example 1.

NFP mutant binding assays: The binding assay using NFP wild type (WT)protein was replicated seven times, while the binding assay using theNFP mutant NFP L147D L154D was replicated four times. A summary ofresults is shown in FIG. 2B.

Biolayer interferometry (BLI): Binding of NFP WT and NFP L147D/L154Dmutant to S. meliloti LCO-IV was measured on an Octet RED 96 system(Pall ForteBio). S. meliloti LCO consists of a tetrameric/pentamericN-acetylglucosamine backbone that is 0-sulfated on the reducing terminalresidue, 0-acetylated on the non-reducing terminal residue, andmono-N-acylated by unsaturated C16 acyl groups. Biotinylated ligandconjugates were immobilized on streptavidin biosensors (kinetic quality,Pall ForteBio) at a concentration of 125-250 nM for 5 minutes. Thebinding assays were replicated 7 times for the NFP WT, and 4 times forthe NFP L147D/L154D mutant. Data analysis was performed in GraphPadPrism 6 software (GraphPad Software, Inc.). Equilibrium dissociationconstants derived from the steady-state were determined by applying anon-linear regression (one site, specific binding) to the response atequilibrium plotted against the protein concentration. Kineticparameters were determined by non-linear regression (associationfollowed by dissociation) on the subtracted data. Results are shown inFIGS. 19A-19C. Binding of A. thaliana CERK1 (AtCERK1) to chitopentaose(CO5) and chitooctaose (C08) was measured in the same way. Results areshown in FIGS. 18A-18B.

Results

FIG. 2A shows modelling of the NFP ectodomain bound to a ligand withpredicted chitin and LCO fatty acid chain locations. Structuralalignment of the NFP ectodomain with ligand-bound CERK1 positions chitinin the LysM2 binding groove of NFP without any obvious clashes.Strikingly, the electrostatic surface potential revealed a hydrophobicpatch on the NFP ectodomain that is located near the non-reducing moietyof the docked chitin molecule, which potentially could accommodate thefatty acid chain of the LCO ligand. Two leucine residues (L147 and L154)were identified as the residues that give this patch its hydrophobiccharacter.

To test the contribution of these two residues to LCO binding, bothresidues were replaced with similarly sized but negatively chargedaspartate residues to produce NFP L147D L154D. Interestingly, the doublemutated NFP L147D L154D ectodomain bound S. meliloti LCO-IV withapproximately two times lower affinity; Kd of 48.0±1.0 μM (FIG. 2B).Closer inspection of the binding kinetics revealed that the association(K_(on)) was almost unaffected whereas the dissociation (K_(off)) wasapproximately 15 times faster in the double mutant. These results showthat the hydrophobic patch of the NFP ectodomain is stabilizing the LCObound state, and that this stabilization is most likely occurring viathe fatty acid chain. Docking the LCO fatty acid in this hydrophobicpatch and the chitin backbone in the LysM2 binding site (derived fromCERK1) would place the sulphate and acetyl side groups facing K141.

Biochemical analysis of LCO binding to the hydrophobic patch mutantreveals that purified L147D/L154D NFP-ECD bound S. meliloti LCO-IV with13-fold lower affinity (Kd of 166.7±4.2 μM) compared to WT NFP-ECD(FIGS. 19A-19C). The association rate (k_(on)) was 4.5-fold faster andthe dissociation rate (k_(off)) was dramatically increased with 59-foldin the double mutant compared to the WT NFP-ECD, suggesting that thehydrophobic patch had a strong stabilizing effect on LCO bindingmediated by the acyl chain.

The binding kinetics of AtCERK1 binding to chitin fragments weremeasured as a comparison. As shown in FIGS. 18A-18B, fast associationand dissociation rates were seen. These kinetics were reminiscent of thekinetics observed for the mutant L147D/L154D NFP-ECD (FIG. 19B). Thebinding kinetics of AtCERK1 to chitin fragments were clearly differentthan the binding kinetics of NFP to LCO (FIG. 19A).

Together, the data provided evidence that the hydrophobic patch in NFP(shown in FIG. 19D) was a conserved structural imprint critical for LCOperception and symbiotic signaling.

Example 3: Complementation Test in Medicago nfp Mutants

To confirm the biochemical observations described in the previousexamples, next a complementation test was performed in Medicago nfpmutants using hairy root transformation.

Materials and Methods

Complementation assay: Construct assembly, plant growth conditions,hairy root transformations, nodulation and ROS assays were generallyconducted as described in Bozsoki et al. (2017) (Bozsoki Z, Cheng J,Feng F, Gysel K, Vinther M, Andersen K R, Oldroyd G, Blaise M, RadutoiuS, Stougaard J (2017) Receptor-mediated chitin perception in legumeroots is functionally separable from Nod factor perception. Proc NatlAcad Sci 114: E8118-E8127). A general schematic of the construct isprovided in FIG. 3. The tested transgenes were the mutated LysMreceptors described in Example 2.

Results

FIGS. 4A-4B shows the results of the complementation test. The resultsshown in FIG. 4A are complementation tests where the plants wereinoculated with S. meliloti strain 2011. When Medicago nfp mutants aretransformed with the wild type Nfp gene, complementation is seen, whichis defined as an average of 10 nodules per plant 6-7 weeks afterinoculation with S. meliloti strain 2011. In contrast, roots transformedwith the double mutant construct (L147D/L154D) did not develop anynodules per plant 6-7 weeks after inoculation with S. meliloti strain2011.

These complementation experiments were repeated using S. medicaeinoculation, which has been reported to nodulate Medicago with higherefficiency. The results shown in FIG. 4B are complementation tests wherethe plants were inoculated with S. medicae. The S. medicae resultsconfirm that the double mutant construct (L147D/L154D) complementspoorly. Taken together, these results show that the hydrophobic patch inNFP is required for LCO recognition, and for functional symbioticsignaling.

Example 4: Functional Characterization of CO and LCO Receptors UsingDomain Swaps

The following example describes functional characterization of the LotusLCO receptor NFR1 and the Lotus CO receptor CERK6. This was done usingdomain swaps, and by measuring nodulation and defense (reactive oxygenspecies, ROS) responses to assess complementation.

Materials and Methods

Complementation assay: Construct assembly, plant growth conditions,hairy root transformations, nodulation and ROS assays were generallyconducted as described in Bozsoki et al. (2017) (Bozsoki Z, Cheng J,Feng F, Gysel K, Vinther M, Andersen K R, Oldroyd G, Blaise M, RadutoiuS, Stougaard J (2017) Receptor-mediated chitin perception in legumeroots is functionally separable from Nod factor perception. Proc NatlAcad Sci 114: E8118-E8127). A general schematic of the construct isprovided in FIG. 3, whereby the pNfr1 promoter was used for theconstructs tested in nodulation assays, and the pCerk6 promoter was usedfor the constructs tested in ROS assays. For functional complementationof nfr1 and cerk6 mutants, only plants expressing the YFP marker proteinfrom the transformation control (FIG. 3) were used. For the nodulationassays, nodules were counted on hairy root transformed L. japonicusnfr1-1 mutant roots after the indicated days post inoculation (dpi)(e.g., 44 dpi, 49 dpi, and 50 dpi) with M. loti R7A (FIG. 5A). For theROS assays, transformed roots were harvested from individual plants,then the root material was divided into two halves, each half beingtested for ROS response to CO8 or flg22. For each transformed plant theratio of CO8 and flg22 elicited ROS peak values after were plottednormalized to the wild type sample, which was set as 1 (FIG. 5B). Thetested chimeric receptors are depicted as shaded block diagrams in FIGS.5A-5B.

Results

FIGS. 5A-5B show results of functional studies measuring nodulation anddefense using domain swaps between the Lotus LCO receptor NFR1 and theLotus CO receptor CERK6. FIG. 5A shows complementation experiments of aLotus nfr1 single mutant with different domain-swapped proteinconstructs expressed under the pNFR1 promoter. Nodulation was used toassess complementation. FIG. 5B shows complementation experiments of aLotus cerk6 single mutant with different domain-swapped proteinconstructs expressed under the pCerk6 promoter. The level of elicitedROS response was used to assess complementation. The results of theseexperiments show that the LysM1 domain of the NFR1 ectodomain isimportant to perceive both LCO (in the case of NFR1) and CO (in the caseof CERK6) ligands.

Additional experiments, also depicted in FIGS. 5A-5B, swapped smallersections, referred to as regions, of the domains. These experimentsshowed that two regions, namely region II and region IV, wereparticularly important for specific recognition of a ligand. Takentogether, these results show that swapping either the entire LysM1domain, or swapping only region II and region IV, is sufficient toconvert a CO receptor into an LCO receptor.

Example 5: Structural Characterization of Lotus CERK6 Ectodomain

The following example describes the structural characterization of theLotus CERK6 protein ectodomain.

Materials and Methods

Modelling: The target LysM receptor amino acid sequence (Lotus CERK6)was aligned with a known receptor sequence (Medicago NFP). Then, theLysM1-3 domains of the target sequence were used as an input inSWISS-MODEL (Biasini 2014). The structural coordinate file (.pdb) of theMedicago NFP crystal structure as template file in SWISS-MODEL (Biasini2014), and the modelling program was run using the command ‘BuildModel’. The electrostatic surface potential of the output target (.pdb)model generated with SWISS-MODEL was calculated using PDB2PQR & APBSwebservers (PMID: 21425296) and visualized in PyMol using APBS tools 2.1(DeLano, W. L. 2002). The 3D structure of the Lotus CERK6 ectodomain isdepicted in FIG. 6, and corresponds to that published by Bozsoki et al.,2017 (Bozsoki Z, Cheng J, Feng F, Gysel K, Vinther M, Andersen K R,Oldroyd G, Blaise M, Radutoiu S, Stougaard J (2017) Receptor-mediatedchitin perception in legume roots is functionally separable from Nodfactor perception. Proc Natl Acad Sci 114: E8118-E8127).

Results

The 3D structure of CERK6 shows that region II and region IV are locatedadjacent to each other as shown in FIG. 6. This indicates the potentialinvolvement of these two regions in a possible binding site.

Example 6: Functional Characterization of LCO Receptors Using DomainSwaps

The following example describes functional characterization of the LotusLCO receptor NFR1 and the Medicago LCO receptor LYK3. This was doneusing domain swaps, and by measuring nodulation to assesscomplementation.

Materials and Methods

Complementation assay: Construct assembly, plant growth conditions,hairy root transformations, nodulation and ROS assays were generallyconducted as described in Bozsoki et al. (2017) (Bozsoki Z, Cheng J,Feng F, Gysel K, Vinther M, Andersen K R, Oldroyd G, Blaise M, RadutoiuS, Stougaard J (2017) Receptor-mediated chitin perception in legumeroots is functionally separable from Nod factor perception. Proc NatlAcad Sci 114: E8118-E8127). A general schematic of the construct isprovided in FIG. 3, whereby the pNFR1 promoter was used to drive thechimeric constructs. The tested chimeric receptors in Lotus are depictedas block diagrams below the graph in FIG. 7, where NFR1 domains areshown in white and LYK3 domains are shown in grey, and transverse linesacross the block depicting the LysM1 domain indicate sections II and IV.Unaltered Lotus CERK6 protein was used as a negative control (zeronodulation). Nodules were counted on hairy root transformed L. japonicusnfr1-1 mutant roots 35 days post inoculation with M. loti R7A. M. lotiR7A is the cognate N-fixing bacterial strain for L. japonicus, and isnot recognized by M. truncatula.

The tested chimeric receptors in Medicago are depicted as block diagramsin FIGS. 17A-17B, where NFR1 domains are shown in black, LYK3 domainsare shown in grey, and transverse lines across the block depicting theLysM1 domain indicate regions II, III, and IV. The pLYK3 promoter wasused to drive the chimeric constructs. Nodules were counted on hairyroot transformed M. truncatula WT or M. truncatula lyk3 mutant roots 35days post inoculation with S. meliloti. S. meliloti is the cognateN-fixing bacterial strain for M. truncatula, and is not recognized by L.japonicus.

Results

FIG. 7 shows results of functional studies measuring nodulation usingdomain swaps between the Lotus LCO receptor NFR1 and the Medicago LCOreceptor LYK3. In addition, LjCERK6 was included as a negative control(zero nodulation). In a Lotus nfr1-1 mutant, two recombinant receptorswere able to complement. Domains of NFR1 (white) and LYK3 (grey)proteins were assembled in different chimeric constructs as shown in thediagram below the graph. Transversal lines across LysM1 show the wherethe section II and IV were derived from. CERK6 protein was used ascontrol. Only chimeric receptors that contained regions II and IV of theLysM1 domain or the entire LysM1 domain from NFR1 complemented the nfr1mutant when inoculated with M. loti. This indicates that the LysM1domain is essential for allowing specific M. loti LCO recognition. Theimportance of region II and region IV is shown by a recombinant receptorentirely consisting of MtLYK3 except for LjNFR1 region hand region IV,which was able to functionally complement the nfr1 mutant, even thoughthe efficiency did not reach wild-type levels. This chimeric MtLYK3receptor shows that the swap of regions II and IV is sufficient tochange the specificity to M. loti LCO. Taken together, these resultsindicate that the LysM1 domain is essential for recognizing those LCOsproduced by the cognate N-fixing bacterial strain of a legume species.Moreover, regions II and IV are particularly important for thisrecognition, because when they are replaced, recognition is lost.

The results from FIG. 7 showed that a chimeric MtLYK3 receptor with aswap of regions II and IV (rightmost receptor on graph) was sufficientto change the specificity of an otherwise fully MtLYK3 protein, whichwould normally recognize S. meliloti LCO, to M. loti LCO. Engineeringthe LjNFR1 receptor with regions II and IV from MtLyk3 resulted in areceptor that was not able to recognize M. loti LCO (receptor third fromleft on graph and receptor third from right on graph). This chimericreceptor showed that the swap of regions II and IV was sufficient toabolish recognition of M. loti by an otherwise fully LjNFR1 protein.

FIG. 17A shows that in a M. truncatula lyk3 mutant, recombinantreceptors containing sections of LysM1 that included region III fromMtLYK3 were able to complement. Region III is six amino acids locatedbetween region II and region IV (FIG. 17B). Engineered receptors thathad the entire LysM1 from MtLYK3 (e.g., the receptor sixth from left(empty vectors counted) or the receptor sixth from right), a section ofLysM1 spanning region II to region IV from MtLYK3 (e.g., the receptorfourth from left (empty vectors counted) or the receptor eighth fromright), or region II, region III, and region IV from MtLYK3 (e.g., thereceptor fifth from left (empty vectors counted) or the receptor seventhfrom right) were all able to complement the symbiotic deficientphenotype of the M. truncatula lyk3 mutant when inoculated with S.meliloti. In contrast, engineered receptors containing only region IIand region IV from MtLYK3 (e.g., the receptor seventh from left (emptyvectors counted) or the receptor fifth from right) were not able tospecifically recognize S. meliloti LCO. It was observed that thechimeric receptor with the MtLYK3 transmembrane and kinase domains(receptor fifth from right) had a very low complementation ability (3plants of the 15 analyzed had 1 or 2 nodules), which was thought to bedue to the high efficiency of these additional regions from MtLYK3. Thisresult was interpreted as region III being required for specific andefficient recognition of S. meliloti LCO, but regions II and IV beingcritical. Engineering the MtLYK3 receptor with regions II to IV fromLjNFR1 (the receptor fourth from right), regions II, III, and IV fromLjNFR1 (the receptor fourth from right), the entire LysM1 from LjNFR1(the receptor second from right), or regions II and IV from LjNFR1 (therightmost receptor) resulted in a receptor that was not able torecognize S. meliloti LCO.

Overall, these results indicated that the LysM1 domain was essential forrecognizing those LCOs produced by the cognate N-fixing bacterial strainof a legume species. When the chimeric receptors were expressed in M.truncatula, the regions II, III, and IV of the LysM1 domain wereidentified as particularly important for this recognition. Replacingregions II and IV were sufficient to obtain a loss of recognition.Replacing regions II, III, and IV were required to obtain gain ofrecognition for S. meliloti LCO and optimal functionality of thereceptor.

Example 7: Engineering Specific LCO Perception

The following example describes engineering of the Lotus receptor LYS11(LjLYS11) to specifically perceive LCOs. This was done using domainswaps, by measuring ligand binding, and by measuring nodulation toassess complementation.

Materials and Methods

LjLYS11 ectodomain production and purification: The LjLYS11 ectodomain(residues 26-234; SEQ ID NO:60) was codon-optimized for insect cellexpression (Genscript, Piscataway, USA) and cloned into the pOET4baculovirus transfer vector (Oxford Expression Technologies). The nativeLjLYS11 signal peptide was replaced with the gp64 signal peptide (SEQ IDNO:59) to facilitate secretion and a hexa-histidine (6×His; SEQ IDNO:61) tag was added to the C-terminus (LAYS11-ecto (26-234), N-termgp64, C-term 6His=SEQ ID NO:56). Recombinant baculoviruses were producedin Sf9 cells (Spodoptera frugiperda) using the FlashBac Gold kit (OxfordExpression technologies) according to the manufacturer's instructionswith Lipofectin (ThermoFisher Scientific) as a transfection reagent.Protein expression was performed as follows. Suspension-cultured Sf9cells were maintained with shaking at 299 K in serum-free MAX-XP(BD-Biosciences, discontinued) or HyClone SFX (GE Healthcare) mediumsupplemented with 1% Pen-Strep (10000 U/ml, Life technologies) and 1% CDlipid concentrate (Gibco). Protein expression was induced by addingrecombinant passage 3 virus once the Sf9 cells reached a cell density of1.0*10{circumflex over ( )}6 cells/ml. After 5-7 days of expression,medium supernatant containing LjLYS11 ectodomains was harvested bycentrifugation. This was followed by an overnight dialysis step against50 mM Tris-HCl pH 8, 200 mM NaCl at 277 K. The LjLYS11 ectodomain wasenriched by two subsequent steps of Ni-IMAC purification (HisTrapexcel/HisTrap HP, both GE Healthcare). For crystallography experiments,N-glycans were removed using the endoglycosidase PNGase F (1:15 (w/w),room temperature, overnight). As a final purification step, LjLYS11ectodomain was purified by SEC on a Superdex 200 10/300 or HiLoadSuperdex 200 16/600 (both GE Healthcare) in phosphate buffered saline atpH 7.2 supplemented to a total of 500 mM NaCl (for binding assays) or 50mM Tris-HCl, 200 mM NaCl (for crystallography).

Biolayer interferometry (BLI): Binding of LjLYS11 ectodomain anddomain-swapped versions of LjLYS11 ectodomain to ligands was measured onan Octet RED 96 system (Pall ForteBio). The ligands used were CO5 chitinoligomer (corresponding to the backbone of S. meliloti LCO-V), M. lotiLCO, and S. meliloti LCO. S. meliloti LCO consists of atetrameric/pentameric N-acetylglucosamine backbone that is O-sulfated onthe reducing terminal residue, 0-acetylated on the non-reducing terminalresidue, and mono-N-acylated by unsaturated C16 acyl groups. M. loti LCOis a pentameric N-acetylglucosamine with a cis-vaccenic acid and acarbamoyl group at the non-reducing terminal residue together with a2,4-O-acetylfucose at the reducing terminal residue. Biotinylated ligandconjugates were immobilized on streptavidin biosensors (kinetic quality,Pall ForteBio) at a concentration of 125-250 nM for 5 minutes. Dataanalysis was performed in GraphPad Prism 6 software (GraphPad Software,Inc.). Equilibrium dissociation constants derived from the steady-statewere determined by applying a non-linear regression (one site, specificbinding) to the response at equilibrium plotted against the proteinconcentration. Kinetic parameters were determined by non-linearregression (association followed by dissociation) on the subtracteddata. The tested chimeric receptors are depicted as block diagrams inFIG. 15B, with LjLYS11 domains shown in black and LjNFR5 and above thebinding assay results in FIGS. 15C-15E.

Complementation assay: The complementation assay was done as in Example6. The tested chimeric receptors are depicted as block diagrams in FIG.15F, where LjNFR5 domains are shown in light grey, LjLYS11 domains areshown in grey, and transverse lines across the block depicting the LysM2domain indicate regions QLGDSYD (SEQ ID NO:63) and GV (SEQ ID NO:64)from LjNFR5. Empty vector and full-length LjLYS11 were used as negativecontrols (zero nodulation). Nodules were counted on hairy roottransformed L. japonicus nfr5-2 mutant roots 35 days post inoculationwith M. loti R7A. M. loti R7A is the cognate N-fixing bacterial strainfor L. japonicus.

Results

Based on modelling and crystal structure determination of LjLYS11ectodomain (FIG. 15A), it was predicted that the receptor would likelybe a LCO receptor. To experimentally validate this prediction, bindingexperiments were performed. As shown in FIG. 15C, LjLYS11 ectodomain wasable to bind CO5 (left graph), M. loti LCO (middle graph; M. loti is thecognate N-fixing bacterial strain for L. japonicus), and S. meliloti LCO(right graph; weak binding). This result indicated that the identifiedhydrophobic patch in the LjLYS11 ectodomain allowed it to bind LCO.Therefore, the hydrophobic patch was predictive of LCO-binding ability.

Next, it was tested whether stringent and specific LCO recognition couldbe engineered. For these tests, LjLYS11 ectodomains were engineered tocontain parts of LjNFR5 receptors. Either the entire LysM2 or keyresidues from the LysM2 hydrophobic patch from LjLYS11 were replacedwith the corresponding regions QLGDSYD (SEQ ID NO:63) and GV (SEQ IDNO:64) from LjNFR5, and ligand binding of these chimeric ectodomains wasmeasured. As shown in FIG. 15D, replacing the entire LysM2 resulted inimproved affinity to LCOs (both M. loti and S. meliloti LCOs), andresulted in a loss of ability to bind CO. A similar result was seen whenonly key residues of LysM2 were replaced (FIG. 15E).

Then, chimeric receptors were tested in planta. For these tests, thesame chimeric LjLYS11 ectodomains were used (the entire LysM2, or keyresidues from LysM2 from LjLYS11 were replaced with the correspondingregions from LjNFR5) or the entire LjLYS11 ectodomain (LysM1, LysM2, andLysM3) was used, and these were attached to the transmembrane domain(wavy shape in schematic of FIG. 15F) and kinase domain (oval shape inschematic of FIG. 15F) of LjNFR5. In addition, full-length LjNFR5 andfull-length LjLYS11 were tested. As shown in FIG. 15F, chimericreceptors with any one of these modifications (the receptors fourth fromright, third from right, and second from right) retained their capacityto perceive the M. loti Nod factor and to initiate a symbiotic signalingevent with similar efficiency as LjNFR5.

Interestingly, the chimeric LjLYS11/LjNFR5 ectodomains had different LCObinding kinetics with slow on/off rates that resembled the bindingkinetics of M. truncatula NFP. As shown in FIG. 18D, slow on/off ratebinding kinetics are thought to be important for functional symbioticsignaling. The fast on/off rate binding kinetics seen with hydrophobicpatch mutants does not result in symbiotic signalling (FIG. 18E).Further, fast on/off kinetics also appear to be a hallmark of COperception (FIG. 18C). As shown in FIG. 18F, NFP shared the cysteinebridge connectivity pattern and the overall arrangement of the scaffoldwith other LysM receptor kinases involved in chitin-elicited defencesignalling. This result supported the hypothesis that despite theirdifferent function, these LysM receptors shared a common evolutionaryorigin (Zhang, X.-C. et al. Molecular evolution of lysin motif-typereceptor-like kinases in plants. Plant Physiol. 144, 623-636 (2007)).The shared structural features of the LysM receptors provided furthersupport for the ability to engineer these receptors to have differentbinding kinetics. For example, the altered binding kinetics observedwith the chimeric LjLYS11/LjNFR5 ectodomains indicate that receptors canbe engineered to have LCO binding kinetics characteristics of functionalsymbiotic signalling.

Taken together, the results seen with chimeric LjLYS11/LjNFR5ectodomains show that LysM2 engineering can create receptors with higherstringency toward LCO as well as higher specificity toward LCO.

Example 8: Identifying Target LysM Receptors for Engineering

The following example describes homology modelling in barley (H.vulgare) to identify target LysM receptors for use in engineering.

Materials and Methods

Modelling: Homology modelling was performed with SWISS-MODEL (Biasini,M. et al. SWISS-MODEL: modelling protein tertiary and quaternarystructure using evolutionary information. Nucleic Acids Res. 42,W252-W258 (2014)). For barley RLK10 (HvRLK10), the crystal structure ofMedicago NFP served as the template model onto which the amino acidsequence of the target receptor was mapped. For barley RLK4 (HvRLK4),the crystal structure of Medicago LYK3 served as the template model ontowhich the amino acid sequence of the target receptor was mapped. Theoutput pdb filled structural model was generated and its electrostaticsurface potential was calculated using the PDB2PQR and APBS webservers(PMID: 21425296). The results were visualized in PyMol using APBS tools2.1 (DeLano, W. L. (2002). PyMOL. DeLano Scientific, San Carlos, Calif.,700).

Expression and purification of ectodomain: The HvRLK10 ectodomain(residues 25-231; SEQ ID NO:66) was codon-optimized for insect cellexpression (Genscript, Piscataway, USA) and cloned into the pOET4baculovirus transfer vector (Oxford Expression Technologies). The nativeHvRLK10 signal peptide was replaced with the gp64 signal peptide (SEQ IDNO:59) to facilitate secretion and a hexa-histidine (6×HIS; SEQ IDNO:61) tag was added to the C-terminus (HvRLK10-ecto (25-231), N-termgp64, C-term 6His=SEQ ID NO:65). The HvRLK4 ectodomain (residues 27-228;SEQ ID NO:68) was codon-optimized for insect cell expression (Genscript,Piscataway, USA) and cloned into the pOET4 baculovirus transfer vector(Oxford Expression Technologies). The native HvRLK4 signal peptide wasreplaced with the gp64 signal peptide (SEQ ID NO:59) to facilitatesecretion and a hexa-histidine (6×HIS; SEQ ID NO:61) tag was added tothe C-terminus (RLK4-ecto (27-228), N-term gp64, C-term 6His=SEQ IDNO:67). Recombinant baculoviruses were produced in Sf9 cells (Spodopterafrugiperda) using the FlashBac Gold kit (Oxford Expression technologies)according to the manufacturer's instructions with Lipofectin(ThermoFisher Scientific) as a transfection reagent.

Protein expression was performed as follows. Suspension-cultured Sf9cells were maintained with shaking at 299 K in serum-free MAX-XP(BD-Biosciences, discontinued) or HyClone SFX (GE Healthcare) mediumsupplemented with 1% Pen-Strep (10000 U/ml, Life technologies) and 1% CDlipid concentrate (Gibco). Protein expression was induced by addingrecombinant passage 3 virus once the Sf9 cells reached a cell density of1.0*10{circumflex over ( )}6 cells/ml. After 5-7 days of expression,medium supernatant containing HvRLK10 ectodomain or HvRLK4 ectodomainwas harvested by centrifugation. This was followed by an overnightdialysis step against 50 mM Tris-HCl pH 8, 200 mM NaCl at 277 K. TheHvRLK10 ectodomain or HvRLK4 ectodomain was enriched by two subsequentsteps of Ni-IMAC purification (HisTrap excel/HisTrap HP, both GEHealthcare).

Biolayer interferometry (BLI): Binding of HvRLK10 ectodomain or HvRLK4ectodomain to ligands was measured on an Octet RED 96 system (PallForteBio). The ligands used were CO5 chitin oligomer (corresponding tothe backbone of S. meliloti LCO-V), M. loti LCO, and S. meliloti LCO. S.meliloti LCO consists of a tetrameric/pentameric N-acetylglucosaminebackbone that is O-sulfated on the reducing terminal residue,0-acetylated on the non-reducing terminal residue, and mono-N-acylatedby unsaturated C16 acyl groups. M. loti LCO is a pentamericN-acetylglucosamine with a cis-vaccenic acid and a carbamoyl group atthe non-reducing terminal residue together with a 2,4-O-acetylfucose atthe reducing terminal residue. Biotinylated ligand conjugates wereimmobilized on streptavidin biosensors (kinetic quality, Pall ForteBio)at a concentration of 125-250 nM for 5 minutes. Data analysis wasperformed in GraphPad Prism 6 software (GraphPad Software, Inc.).Equilibrium dissociation constants derived from the steady-state weredetermined by applying a non-linear regression (one site, specificbinding) to the response at equilibrium plotted against the proteinconcentration. Kinetic parameters were determined by non-linearregression (association followed by dissociation) on the subtracteddata.

Results

Homology modelling of all ten barley LysM receptor-like kinases (RLKs)was done using the Medicago NFP structure as a template. Of the barleyLysM RLKs, HvRLK10 was the receptor that was closest to Medicago NFP andmodelled the best using this approach. FIG. 16B shows homology modellingresults for HvRLK10, which revealed that the hydrophobic patch wasindeed present in the equivalent positions immediately below the LysM2domain of this receptor. This clear hydrophobic patch indicated thatHvRLK10 was a NFP/NFR5 type of LCO receptor.

To experimentally validate this prediction, the HvRLK10 ectodomain wasexpressed and purified for use in binding experiments (ectodomainschematic shown at top of FIG. 16A). The HvRLK10 ectodomain was shown tobind both M. loti LCO (FIG. 16C) and S. meliloti LCO (FIG. 16D). Incontrast, the HvRLK10 ectodomain did not bind CO5 (FIG. 16A). Theseresults provided functional characterization of the HvRLK10 ectodomain,and showed that it bound LCOs, but not COs. The results confirmed thatthe HvRLK10 receptor was a LCO receptor, as had been predicted by thehomology modelling.

In addition, homology modelling of all ten barley LysM RLKs was doneusing the Medicago LYK3 structure as a template. HvRLK4 was the receptorthat was closest to Medicago LYK3 and modelled the best using thisapproach. FIG. 16F shows homology modelling results for HvRLK4. Theseresults indicated that this receptor was a NFR1/LYK3 type of LCOreceptor.

To experimentally validate this prediction, the HvRLK4 ectodomain wasexpressed and purified for use in binding experiments (ectodomainschematic shown at top of FIG. 16E). The HvRLK4 ectodomain was shown tobind both M. loti LCO (FIG. 16G) and S. meliloti LCO (FIG. 16H). Incontrast, the HvRLK4 ectodomain did not bind CO5 (FIG. 16E). Theseresults provided functional characterization of the HvRLK4 ectodomain,and showed that it bound LCOs, but not COs. The results confirmed thatthe HvRLK4 receptor was a LCO receptor, as had been predicted by thehomology modelling.

Both HvRLK10 and HvRLK4 were initially identified by homology modelling,and then confirmed to be LCO receptors by biochemical characterization.HvRLK10 and HvRLK4 therefore represent promising target receptors forengineering in barley particularly for engineering receptors thatrecognize LCOs in a manner similar to the donor receptors used to selectthem, Medicago NFP and Medicago LYK3, respectively.

Overall, these results show that the homology modelling approach can beused to identify LCO receptors of both the NFP/NFR5 type and theNFR1/LYK3 type specifically and that this method may be used to identifygood target LysM receptor to modify to alter a desired receptorcharacteristic to be that of the donor LysM receptor used to select thetarget LysM receptor.

Example 9: Exemplary Structural Alignment to Identify of Target Residuesto Modify for Insertion of a Hydrophobic Patch

One of skill in the art would have no difficulty applying the teachingsof this disclosure to genetically alter LysM receptors to include ahydrophobic patch or alter an existing hydrophobic patch. Exemplarysteps would be:

1. Align the target LysM receptor amino acid sequence with one or moreknown LCO receptor sequences (See, e.g., FIGS. 8A-8C, FIGS. 9A-9B, FIGS.10A-10B, FIGS. 11A-11B) to identify the sequence of the LysM1-3 domainsin the target amino acid sequence.

Applying this step to the HvLysM-RLK2/37-247 sequence produced thefollowing amino acid sequence:

>HvLySM-RLK2/37-247 SVEGFNCSANGTYPCQAYALYRAGLAGVPPDLSAAGDLFGVSRFMLAHANNLSTSAAPAAGQ PLLVPLQCGCPSGSPNAYAPTQYQISSGDTFWIVSVTKLQNLTQYQAVERVNPTVVPTKLEV GDMVTFPIFCQCPTAAQNATALVTYVMQQGDTYASIAAAFAVDAQSLVSLNGPEQGTQLFSE ILVPLRRQVPKWLPPIVTRNDASAT2. Use the LysM1-3 domain amino acid sequence as the input sequence tobe modeled in an appropriate molecular modeling program such asSWISS-MODEL (Biasini 2014). SWISS-MODEL can be readily accessed atswissmodel.expasy.org under interactive#structure.3. Input the structural template to the molecular modelling program, forexample from a structural coordinate file (e.g., a pdb format file).

The HvLysM-RLK2/37-247 LysM1-3 domain amino acid sequence was enteredinto SWISS-MODEL as was the Medicago NFP receptor ectodomain crystalstructure .pdb file (the atomic coordinates are reproduced at the end ofthe specification). The SWISS-MODEL program was run by the command‘Build Model’. The Medicago NFP receptor ectodomain crystal structurewas chosen as it has a known hydrophobic patch. One of skill in the artcan readily select others based upon the teachings in thisspecification.

4. Optionally create an electrostatic surface potential of the targetmodel and structurally align with a structure with chitin (or glycan)bound to the LysM2 domain to align the ligand binding grooves.

An electrostatic surface potential of the output target (.pdb) modelgenerated with SWISS-MODEL was calculated using PDB2PQR & APBSwebservers (PMID: 21425296) and visualized in PyMol using APBS tools 2.1(DeLano, W. L. 2002). The AtCERK1 ectodomain structure (PDB coordinates4EBZ) which has the chitin bound in the structure was aligned to thetarget model in PyMol. One of skill in the art would readily understandthe position of the chitin binding domain as the LysM chitin bindingmotif is defined structurally in Liu et al. Science 2012 for AtCERK1.This aligned the chitin (C04) ligand in the LysM2 ligand binding grooveof the target model. FIGS. 13A-13B show the PyMol visualization of theLysM1-3 domains of the HvLysM-RLK2/37-247 model with the LysM1, LysM2,and LysM3 domains labeled (FIG. 13A), and the electrostatic surfacepotential of the model with chitin modeled in the binding groove (FIG.13B).

5. Select the residues from the alignment in the target model that alignwith the known hydrophobic patch.

From the sequence alignment (1), structural alignment of the targetmodel with the crystal structure of Medicago NFP and the electrostaticsurface potential information (5) the hydrophobic patch was identified(with the placed chitin from AtCERK1 as reference for locating the CObinding groove as shown in (FIG. 13B). Hot-spot residues correspondingto the Medicago NFP ectodomain hydrophobic patch (L147, L151, L152,L154, T156, K157 and V158) were identified based on the amino acid beingwithin 3 Å of an alpha carbon of a known hydrophobic patch amino acidresidue (Medicago NFP L147, L151, L152, L154, T156, K157 and V158) inthe structural alignment. As one of skill in the art would appreciate,residues like lysine (K) and arginine (R) that are not classicallycharacterized as hydrophobic, do contain hydrophobic properties relatedto the Calpha, Cbeta, Cgamma, Cdelta and Cepsilon atoms that might beimportant for LCO binding, selectivity, promiscuity, stringency, andaffinity and therefore are still potentially important (e.g., K157 ofthe Medicago NFP hydrophobic patch). The identified residues in theHvLysM-RLK2/37-247 model (bolded in FIG. 13C) can be mutated, preferablywith additional modeling, to obtain engineered LCO binding, LCO/COselectivity, LCO promiscuity, LCO stringency, LCO affinity. One of skillin the art would appreciate that similar structural modeling can be usedto structurally align LysM1 domains to identify regions II and IV inorder to substitute and alter specificity, affinity and selectivity of atarget LysM receptor for an agonist.

The Medicago NFP ectodomain crystal Structure LINK C1 NAG A2076 O4 NAG A1076 LINK C1 NAG A2123 O4 NAG A 1123 LINK C1 NAG A2144 O4 NAG A 1144LINK C1 NAG A2228 O4 NAG A 1228 LINK C1 NAG A1076 ND2 ASN A 76 LINK C1NAG A1123 ND2 ASN A 123 LINK C1 NAG A1144 ND2 ASN A 144 LINK C1 NAGA1228 ND2 ASN A 228 SSBOND 1 CYS A 50 CYS A 115 SSBOND 2 CYS A 58 CYS A177 SSBOND 3 CYS A 113 CYS A 175 CRYST1 77.410 98.160 71.890 90.00 90.0090.00 C 2 2 21 SCALE1 0.012918 0.000000 0.000000 0.00000 SCALE2 0.0000000.010187 0.000000 0.00000 SCALE3 0.000000 0.000000 0.013910 0.00000 ATOM1 N SER A 44 47.490 34.635 22.387 1.00 132.78 A N ANISOU 1 N SER A 4414506 18967 16976 −687 −2436 7410 A N ATOM 2 CA SER A 44 48.575 35.49722.832 1.00 128.50 A C ANISOU 2 CA SER A 44 14079 17864 16882 −714 −25777389 A C ATOM 3 C SER A 44 49.218 34.959 24.094 1.00 121.82 A C ANISOU 3C SER A 44 13323 16878 16087 −720 −2520 7111 A C ATOM 4 O SER A 4450.272 34.341 24.053 1.00 114.50 A O ANISOU 4 O SER A 44 12311 1624114951 −801 −2396 7114 A O ATOM 5 CB SER A 44 48.060 36.911 23.094 1.00126.83 A C ANISOU 5 CB SER A 44 13992 17346 16852 −547 −2697 7268 A CATOM 6 OG SER A 44 49.010 37.665 23.828 1.00 128.24 A O ANISOU 6 OG SERA 44 14237 17006 17480 −596 −2848 7359 A O ATOM 7 N GLU A 45 48.55135.184 25.216 1.00 120.97 A N ANISOU 7 N GLU A 45 13377 16340 16246 −626−2607 6862 A N ATOM 8 CA GLU A 45 49.040 34.751 26.516 1.00 122.62 A CANISOU 8 CA GLU A 45 13672 16343 16575 −651 −2584 6624 A C ATOM 9 C GLUA 45 49.060 33.238 26.635 1.00 120.68 A C ANISOU 9 C GLU A 45 1345816423 15972 −535 −2435 6316 A C ATOM 10 O GLU A 45 48.274 32.543 25.9981.00 122.15 A O ANISOU 10 O GLU A 45 13634 16922 15853 −411 −2368 6224 AO ATOM 11 CB GLU A 45 48.163 35.326 27.628 1.00 121.51 A C ANISOU 11 CBGLU A 45 13688 15639 16840 −586 −2726 6442 A C ATOM 12 CG GLU A 4548.212 36.836 27.755 1.00 127.27 A C ANISOU 12 CG GLU A 45 14403 1599817957 −715 −2869 6708 A C ATOM 13 CD GLU A 45 49.555 37.327 28.243 1.00132.32 A C ANISOU 13 CD GLU A 45 14930 16696 18650 −952 −2852 6959 A CATOM 14 OE1 GLU A 45 49.586 38.123 29.205 1.00 136.87 A O ANISOU 14 OE1GLU A 45 15373 17434 19196 −1069 −2872 7310 A O ATOM 15 OE2 GLU A 4550.579 36.907 27.669 1.00 131.61 A O1− ANISOU 15 OE2 GLU A 45 1487716507 18623 −1022 −2821 6805 A O1− ATOM 16 N THR A 46 49.991 32.74327.440 1.00 124.90 A N ANISOU 16 N THR A 46 14032 16873 16550 −581 −23896151 A N ATOM 17 CA THR A 46 50.130 31.318 27.669 1.00 110.09 A C ANISOU17 CA THR A 46 12173 15318 14337 −492 −2238 5885 A C ATOM 18 C THR A 4649.107 30.887 28.697 1.00 103.70 A C ANISOU 18 C THR A 46 11505 1443713459 −283 −2255 5524 A C ATOM 19 O THR A 46 49.388 30.892 29.895 1.00100.49 A O ANISOU 19 O THR A 46 11098 14391 12694 −175 −2139 5343 A OATOM 20 CB THR A 46 51.526 30.968 28.209 1.00 90.86 A C ANISOU 20 CB THRA 46 9735 12798 11989 −599 −2195 5824 A C ATOM 21 OG1 THR A 46 51.55029.598 28.624 1.00 82.38 A O ANISOU 21 OG1 THR A 46 8675 12043 10583−499 −2045 5566 A O ATOM 22 CG2 THR A 46 51.868 31.850 29.398 1.00 77.97A C ANISOU 22 CG2 THR A 46 8237 10610 10779 −617 −2337 5681 A C ATOM 23N ASN A 47 47.911 30.561 28.230 1.00 89.04 A N ANISOU 23 N ASN A 47 976512123 11942 −226 −2396 5399 A N ATOM 24 CA ASN A 47 46.844 30.100 29.1041.00 83.06 A C ANISOU 24 CA ASN A 47 9114 11294 11149 −31 −2436 5105 A CATOM 25 C ASN A 47 46.404 31.034 30.221 1.00 80.85 A C ANISOU 25 C ASN A47 8941 10474 11302 9 −2591 5004 A C ATOM 26 O ASN A 47 46.004 32.16129.974 1.00 86.84 A O ANISOU 26 O ASN A 47 9724 11126 12145 112 −26715008 A O ATOM 27 CB ASN A 47 47.083 28.674 29.595 1.00 84.42 A C ANISOU27 CB ASN A 47 9345 11671 11058 72 −2332 4752 A C ATOM 28 CG ASN A 4746.851 27.659 28.508 1.00 86.04 A C ANISOU 28 CG ASN A 47 9512 1236010818 171 −2218 4673 A C ATOM 29 OD1 ASN A 47 46.815 26.461 28.751 1.0087.76 A O ANISOU 29 OD1 ASN A 47 9702 12671 10972 219 −2256 4778 A OATOM 30 ND2 ASN A 47 46.678 28.143 27.293 1.00 88.00 A N ANISOU 30 ND2ASN A 47 9761 12911 10762 198 −2077 4483 A N ATOM 31 N PHE A 48 46.49830.555 31.453 1.00 77.89 A N ANISOU 31 N PHE A 48 8628 9762 11206 −71−2633 4906 A N ATOM 32 CA PHE A 48 45.991 31.292 32.602 1.00 75.80 A CANISOU 32 CA PHE A 48 8491 9027 11281 11 −2743 4640 A C ATOM 33 C PHE A48 46.493 32.702 32.781 1.00 86.98 A C ANISOU 33 C PHE A 48 9926 1001813104 −64 −2869 4809 A C ATOM 34 O PHE A 48 47.684 32.980 32.841 1.0094.16 A O ANISOU 34 O PHE A 48 10808 10921 14046 −4 −2923 4965 A O ATOM35 CB PHE A 48 46.320 30.503 33.864 1.00 65.86 A C ANISOU 35 CB PHE A 487303 7645 10076 −14 −2713 4362 A C ATOM 36 CG PHE A 48 46.172 29.03433.683 1.00 63.01 A C ANISOU 36 CG PHE A 48 6933 7684 9323 87 −2597 4158A C ATOM 37 CD2 PHE A 48 47.112 28.323 32.976 1.00 62.96 A C ANISOU 37CD2 PHE A 48 6835 8073 9013 6 −2469 4295 A C ATOM 38 CD1 PHE A 48 45.06628.381 34.164 1.00 60.68 A C ANISOU 38 CD1 PHE A 48 6713 7401 8941 267−2609 3827 A C ATOM 39 CE2 PHE A 48 46.969 26.974 32.779 1.00 60.62 A CANISOU 39 CE2 PHE A 48 6536 8154 8341 108 −2350 4087 A C ATOM 40 CE1 PHEA 48 44.914 27.031 33.971 1.00 58.32 A C ANISOU 40 CE1 PHE A 48 64137479 8266 348 −2488 3607 A C ATOM 41 CZ PHE A 48 45.864 26.329 33.2741.00 58.33 A C ANISOU 41 CZ PHE A 48 6338 7852 7972 270 −2349 3723 A CATOM 42 N THR A 49 45.510 33.574 32.909 1.00 82.31 A N ANISOU 42 N THR A49 9387 9064 12823 −188 −2920 4756 A N ATOM 43 CA THR A 49 45.676 34.99233.123 1.00 98.91 A C ANISOU 43 CA THR A 49 11496 10792 15293 −304 −30294935 A C ATOM 44 C THR A 49 44.436 35.410 33.889 1.00 99.11 A C ANISOU44 C THR A 49 11606 10480 15570 −157 −3127 4794 A C ATOM 45 O THR A 4943.357 34.846 33.711 1.00 96.68 A O ANISOU 45 O THR A 49 11283 996115491 −209 −3212 5006 A O ATOM 46 CB THR A 49 45.709 35.771 31.797 1.00109.76 A C ANISOU 46 CB THR A 49 12731 12391 16580 −449 −3025 5389 A CATOM 47 OG1 THR A 49 44.666 35.294 30.938 1.00 106.67 A O ANISOU 47 OG1THR A 49 12281 12270 15978 −332 −3012 5538 A O ATOM 48 CG2 THR A 4947.041 35.602 31.095 1.00 111.16 A C ANISOU 48 CG2 THR A 49 12807 1292116508 −598 −2915 5529 A C ATOM 49 N CYS A 50 44.549 36.431 34.711 1.0092.80 A N ANISOU 49 N CYS A 50 10892 9630 14739 27 −3116 4435 A N ATOM50 CA CYS A 50 43.388 36.843 35.468 1.00 92.80 A C ANISOU 50 CA CYS A 5010978 9292 14991 171 −3192 4217 A C ATOM 51 C CYS A 50 42.305 37.43134.561 1.00 97.62 A C ANISOU 51 C CYS A 50 11539 9963 15591 278 −32434441 A C ATOM 52 O CYS A 50 42.612 38.040 33.542 1.00 93.35 A O ANISOU52 O CYS A 50 10906 9567 14996 182 −3258 4821 A O ATOM 53 CB CYS A 5043.807 37.863 36.515 1.00 100.59 A C ANISOU 53 CB CYS A 50 12040 980816371 64 −3262 4119 A C ATOM 54 SG CYS A 50 42.820 37.796 38.012 1.00113.19 A S ANISOU 54 SG CYS A 50 13708 11276 18022 −52 −3216 3794 A SATOM 55 N PRO A 51 41.028 37.208 34.907 1.00 111.76 A N ANISOU 55 N PROA 51 13379 11653 17432 474 −3270 4214 A N ATOM 56 CA PRO A 51 39.91137.793 34.166 1.00 113.17 A C ANISOU 56 CA PRO A 51 13516 11829 17655585 −3332 4416 A C ATOM 57 C PRO A 51 39.756 39.242 34.592 1.00 105.69 AC ANISOU 57 C PRO A 51 12623 10423 17113 568 −3426 4447 A C ATOM 58 OPRO A 51 39.514 39.517 35.764 1.00 98.68 A O ANISOU 58 O PRO A 51 118319213 16451 618 −3441 4121 A O ATOM 59 CB PRO A 51 38.718 36.977 34.6481.00 118.11 A C ANISOU 59 CB PRO A 51 14160 12600 18115 806 −3308 4130 AC ATOM 60 CG PRO A 51 39.086 36.599 36.038 1.00 118.64 A C ANISOU 60 CGPRO A 51 14325 12479 18272 816 −3271 3702 A C ATOM 61 CD PRO A 51 40.56836.342 36.004 1.00 117.49 A C ANISOU 61 CD PRO A 51 14183 12346 18114611 −3232 3770 A C ATOM 62 N VAL A 52 39.897 40.159 33.645 1.00 116.60 AN ANISOU 62 N VAL A 52 13941 11785 18577 498 −3487 4833 A N ATOM 63 CAVAL A 52 39.805 41.578 33.949 1.00 117.67 A C ANISOU 63 CA VAL A 5214126 11488 19094 454 −3580 4905 A C ATOM 64 C VAL A 52 38.391 42.13733.940 1.00 122.26 A C ANISOU 64 C VAL A 52 14743 11887 19825 673 −36364811 A C ATOM 65 O VAL A 52 38.127 43.157 34.571 1.00 120.01 A O ANISOU65 O VAL A 52 14537 11203 19857 700 −3691 4683 A O ATOM 66 CB VAL A 5240.659 42.406 32.975 1.00 116.82 A C ANISOU 66 CB VAL A 52 13934 1143219022 267 −3627 5366 A C ATOM 67 CG1 VAL A 52 40.126 43.825 32.878 1.00118.92 A C ANISOU 67 CG1 VAL A 52 14256 11249 19680 218 −3730 5446 A CATOM 68 CG2 VAL A 52 42.116 42.396 33.409 1.00 109.61 A C ANISOU 68 CG2VAL A 52 12973 10720 17953 53 −3562 5448 A C ATOM 69 N ASP A 53 37.47841.484 33.230 1.00 118.46 A N ANISOU 69 N ASP A 53 14200 11702 19106 835−3620 4860 A N ATOM 70 CA ASP A 53 36.112 42.001 33.169 1.00 124.86 A CANISOU 70 CA ASP A 53 15021 12394 20024 1052 −3671 4808 A C ATOM 71 CASP A 53 35.126 41.211 34.021 1.00 117.67 A C ANISOU 71 C ASP A 53 1416211498 19050 1236 −3622 4372 A C ATOM 72 O ASP A 53 33.916 41.293 33.7941.00 110.53 A O ANISOU 72 O ASP A 53 13233 10639 18122 1433 −3644 4330 AO ATOM 73 CB ASP A 53 35.624 42.040 31.724 1.00 139.47 A C ANISOU 73 CBASP A 53 16759 14568 21665 1114 −3699 5161 A C ATOM 74 CG ASP A 5336.545 42.840 30.834 1.00 155.01 A C ANISOU 74 CG ASP A 53 18664 1654623688 929 −3746 5602 A C ATOM 75 OD1 ASP A 53 37.265 43.718 31.374 1.00159.96 A O ANISOU 75 OD1 ASP A 53 19345 16824 24608 799 −3791 5646 A OATOM 76 OD2 ASP A 53 36.550 42.588 29.603 1.00 160.52 A O1− ANISOU 76OD2 ASP A 53 19253 17611 24126 906 −3738 5895 A O1− ATOM 77 N SER A 5435.610 40.452 34.990 1.00 120.11 A N ANISOU 77 N SER A 54 14531 1177919326 1172 −3556 4053 A N ATOM 78 CA SER A 54 34.790 39.596 35.836 1.00123.38 A C ANISOU 78 CA SER A 54 14987 12234 19659 1316 −3500 3629 A CATOM 79 C SER A 54 35.259 39.713 37.272 1.00 125.41 A C ANISOU 79 C SERA 54 15348 12172 20129 1235 −3474 3284 A C ATOM 80 O SER A 54 36.31940.283 37.552 1.00 125.91 A O ANISOU 80 O SER A 54 15445 12038 203571058 −3494 3378 A O ATOM 81 CB SER A 54 34.856 38.128 35.376 1.00 119.00A C ANISOU 81 CB SER A 54 14380 12123 18712 1326 −3429 3574 A C ATOM 82OG SER A 54 34.328 37.982 34.070 1.00 118.74 A O ANISOU 82 OG SER A 5414247 12423 18445 1404 −3447 3853 A O ATOM 83 N PRO A 55 34.482 39.18938.219 1.00 130.50 A N ANISOU 83 N PRO A 55 16040 12781 20764 1353 −34282874 A N ATOM 84 CA PRO A 55 34.996 39.044 39.577 1.00 129.39 A C ANISOU84 CA PRO A 55 15988 12434 20741 1257 −3383 2517 A C ATOM 85 C PRO A 5536.197 38.126 39.585 1.00 123.67 A C ANISOU 85 C PRO A 55 15261 1188419842 1090 −3336 2530 A C ATOM 86 O PRO A 55 36.288 37.184 38.775 1.00126.40 A O ANISOU 86 O PRO A 55 15546 12573 19906 1106 −3308 2660 A OATOM 87 CB PRO A 55 33.813 38.431 40.342 1.00 128.42 A C ANISOU 87 CBPRO A 55 15885 12361 20548 1424 −3330 2112 A C ATOM 88 CG PRO A 5532.901 37.892 39.286 1.00 128.89 A C ANISOU 88 CG PRO A 55 15858 1273620380 1582 −3339 2264 A C ATOM 89 CD PRO A 55 33.062 38.817 38.131 1.00130.49 A C ANISOU 89 CD PRO A 55 16004 12921 20654 1573 −3416 2719 A CATOM 90 N PRO A 56 37.157 38.366 40.475 1.00 118.72 A N ANISOU 90 N PROA 56 14695 11054 19359 929 −3325 2403 A N ATOM 91 CA PRO A 56 38.40037.588 40.446 1.00 108.59 A C ANISOU 91 CA PRO A 56 13402 9928 17929 763−3286 2460 A C ATOM 92 C PRO A 56 38.343 36.302 41.258 1.00 102.85 A CANISOU 92 C PRO A 56 12713 9342 17023 789 −3204 2076 A C ATOM 93 O PRO A56 39.000 35.326 40.882 1.00 101.03 A O ANISOU 93 O PRO A 56 12454 936016571 734 −3165 2148 A O ATOM 94 CB PRO A 56 39.426 38.573 41.013 1.00106.15 A C ANISOU 94 CB PRO A 56 13131 9329 17871 577 −3322 2519 A CATOM 95 CG PRO A 56 38.631 39.431 41.954 1.00 108.06 A C ANISOU 95 CGPRO A 56 13441 9272 18345 663 −3339 2249 A C ATOM 96 CD PRO A 56 37.22239.496 41.421 1.00 113.49 A C ANISOU 96 CD PRO A 56 14102 10022 18998884 −3355 2260 A C ATOM 97 N SER A 57 37.572 36.266 42.350 1.00 94.60 AN ANISOU 97 N SER A 57 11727 8168 16048 871 −3171 1675 A N ATOM 98 CASER A 57 37.539 35.116 43.241 1.00 87.59 A C ANISOU 98 CA SER A 57 108797404 15000 875 −3089 1290 A C ATOM 99 C SER A 57 36.227 34.364 43.0731.00 79.26 A C ANISOU 99 C SER A 57 9804 6543 13768 1064 −3061 1108 A CATOM 100 O SER A 57 35.162 34.981 42.975 1.00 89.94 A O ANISOU 100 O SERA 57 11141 7819 15214 1193 −3089 1097 A O ATOM 101 CB SER A 57 37.71035.542 44.692 1.00 91.40 A C ANISOU 101 CB SER A 57 11426 7674 15629 798−3052 955 A C ATOM 102 OG SER A 57 38.093 34.435 45.490 1.00 92.41 A OANISOU 102 OG SER A 57 11581 7949 15581 742 −2969 660 A O ATOM 103 N CYSA 58 36.357 33.055 42.866 1.00 68.90 A N ANISOU 103 N CYS A 58 8492 548712201 1080 −3006 961 A N ATOM 104 CA CYS A 58 35.226 32.186 42.590 1.0072.63 A C ANISOU 104 CA CYS A 58 8925 6227 12445 1243 −2991 883 A C ATOM105 C CYS A 58 35.514 30.734 42.965 1.00 61.26 A C ANISOU 105 C CYS A 587515 5006 10756 1224 −2921 627 A C ATOM 106 O CYS A 58 36.592 30.41843.450 1.00 56.36 A O ANISOU 106 O CYS A 58 6931 4354 10129 1096 −2892599 A O ATOM 107 CB CYS A 58 34.888 32.277 41.093 1.00 85.20 A C ANISOU107 CB CYS A 58 10428 8024 13921 1304 −3045 1309 A C ATOM 108 SG CYS A58 35.994 31.399 39.947 1.00 103.73 A S ANISOU 108 SG CYS A 58 1275110337 16325 1115 −3076 1740 A S ATOM 109 N GLU A 59 34.539 29.858 42.7421.00 60.93 A N ANISOU 109 N GLU A 59 7452 5194 10503 1352 −2896 445 A NATOM 110 CA GLU A 59 34.692 28.438 43.035 1.00 52.11 A C ANISOU 110 CAGLU A 59 6368 4304 9129 1349 −2831 184 A C ATOM 111 C GLU A 59 34.63327.639 41.738 1.00 47.29 A C ANISOU 111 C GLU A 59 5684 4068 8215 1420−2850 397 A C ATOM 112 O GLU A 59 33.701 27.807 40.946 1.00 52.46 A OANISOU 112 O GLU A 59 6268 4876 8788 1525 −2886 529 A O ATOM 113 CB GLUA 59 33.606 27.967 44.008 1.00 46.43 A C ANISOU 113 CB GLU A 59 56853598 8360 1403 −2773 −244 A C ATOM 114 CG GLU A 59 33.389 28.875 45.2411.00 60.21 A C ANISOU 114 CG GLU A 59 7464 5073 10340 1334 −2749 −438 AC ATOM 115 CD GLU A 59 34.557 28.856 46.271 1.00 75.79 A C ANISOU 115 CDGLU A 59 9490 6941 12367 1157 −2689 −564 A C ATOM 116 OE1 GLU A 5935.364 27.889 46.275 1.00 76.96 A O ANISOU 116 OE1 GLU A 59 9668 721912355 1097 −2646 −613 A O ATOM 117 OE2 GLU A 59 34.659 29.813 47.0911.00 79.30 A O1− ANISOU 117 OE2 GLU A 59 9937 7199 12994 1089 −2680 −609A O1− ATOM 118 N THR A 60 35.623 26.779 41.521 1.00 45.71 A N ANISOU 118N THR A 60 5493 4048 7825 1355 −2822 430 A N ATOM 119 CA THR A 60 35.73126.019 40.266 1.00 52.03 A C ANISOU 119 CA THR A 60 6231 5286 8252 1352−2770 620 A C ATOM 120 C THR A 60 36.275 24.623 40.587 1.00 42.81 A CANISOU 120 C THR A 60 5128 4362 6776 1277 −2612 345 A C ATOM 121 O THR A60 36.201 24.161 41.730 1.00 40.64 A O ANISOU 121 O THR A 60 4934 39526555 1266 −2565 −14 A O ATOM 122 CB THR A 60 36.581 26.793 39.229 1.0048.35 A C ANISOU 122 CB THR A 60 5689 4858 7824 1278 −2816 1111 A C ATOM123 OG1 THR A 60 36.518 26.138 37.954 1.00 48.75 A O ANISOU 123 OG1 THRA 60 5668 5355 7499 1287 −2764 1294 A O ATOM 124 CG2 THR A 60 38.02426.915 39.669 1.00 47.96 A C ANISOU 124 CG2 THR A 60 5677 4680 7865 1117−2761 1178 A C ATOM 125 N TYR A 61 36.793 23.927 39.578 1.00 42.76 A NANISOU 125 N TYR A 61 5085 4722 6440 1233 −2528 506 A N ATOM 126 CA TYRA 61 37.179 22.539 39.764 1.00 40.22 A C ANISOU 126 CA TYR A 61 48264648 5808 1190 −2378 243 A C ATOM 127 C TYR A 61 38.529 22.289 39.1101.00 40.64 A C ANISOU 127 C TYR A 61 4849 4896 5696 1087 −2289 482 A CATOM 128 O TYR A 61 38.967 23.032 38.240 1.00 43.93 A O ANISOU 128 O TYRA 61 5181 5363 6147 1056 −2338 866 A O ATOM 129 CB TYR A 61 36.12421.571 39.203 1.00 39.50 A C ANISOU 129 CB TYR A 61 4734 4882 5394 1272−2342 75 A C ATOM 130 CG TYR A 61 34.859 21.547 40.017 1.00 42.21 A CANISOU 130 CG TYR A 61 5109 5074 5854 1360 −2400 −233 A C ATOM 131 CD1TYR A 61 33.833 22.463 39.791 1.00 48.15 A C ANISOU 131 CD1 TYR A 615789 5717 6788 1464 −2533 −114 A C ATOM 132 CD2 TYR A 61 34.690 20.61941.019 1.00 43.46 A C ANISOU 132 CD2 TYR A 61 5362 5206 5945 1343 −2321−634 A C ATOM 133 CE1 TYR A 61 32.667 22.448 40.565 1.00 50.86 A CANISOU 133 CE1 TYR A 61 6149 5938 7239 1554 −2580 −403 A C ATOM 134 CE2TYR A 61 33.542 20.594 41.797 1.00 42.89 A C ANISOU 134 CE2 TYR A 615309 5015 5972 1416 −2369 −917 A C ATOM 135 CZ TYR A 61 32.535 21.49741.581 1.00 48.98 A C ANISOU 135 CZ TYR A 61 6003 5690 6918 1523 −2494−811 A C ATOM 136 OH TYR A 61 31.418 21.414 42.405 1.00 45.79 A O ANISOU136 OH TYR A 61 5609 5189 6600 1599 −2529 −1112 A O ATOM 137 N VAL A 6239.182 21.228 39.552 1.00 38.47 A N ANISOU 137 N VAL A 62 4641 4735 52421038 −2156 254 A N ATOM 138 CA VAL A 62 40.508 20.850 39.098 1.00 38.61A C ANISOU 138 CA VAL A 62 4633 4942 5096 953 −2050 421 A C ATOM 139 CVAL A 62 40.444 19.365 38.774 1.00 43.15 A C ANISOU 139 C VAL A 62 52575854 5282 986 −1909 191 A C ATOM 140 O VAL A 62 39.796 18.599 39.4941.00 42.73 A O ANISOU 140 O VAL A 62 5294 5762 5178 1025 −1878 −168 A OATOM 141 CB VAL A 62 41.567 21.143 40.184 1.00 37.82 A C ANISOU 141 CBVAL A 62 4566 4571 5233 852 −2033 371 A C ATOM 142 CG1 VAL A 62 42.60020.052 40.238 1.00 36.37 A C ANISOU 142 CG1 VAL A 62 4406 4604 4808 806−1876 275 A C ATOM 143 CG2 VAL A 62 42.195 22.513 39.992 1.00 40.26 A CANISOU 143 CG2 VAL A 62 4800 4674 5823 772 −2134 742 A C ATOM 144 N ALAA 63 41.084 18.943 37.689 1.00 37.91 A N ANISOU 144 N ALA A 63 4540 55234340 971 −1824 389 A N ATOM 145 CA ALA A 63 40.997 17.540 37.298 1.0036.64 A C ANISOU 145 CA ALA A 63 4436 5679 3807 1009 −1691 167 A C ATOM146 C ALA A 63 42.219 16.762 37.774 1.00 35.29 A C ANISOU 146 C ALA A 634308 5545 3554 965 −1551 55 A C ATOM 147 O ALA A 63 43.356 17.193 37.5801.00 36.35 A O ANISOU 147 O ALA A 63 4370 5696 3743 903 −1522 305 A OATOM 148 CB ALA A 63 40.833 17.397 35.785 1.00 38.60 A C ANISOU 148 CBALA A 63 4606 6310 3750 1036 −1670 399 A C ATOM 149 N TYR A 64 41.97515.599 38.362 1.00 33.11 A N ANISOU 149 N TYR A 64 4144 5295 3141 997−1464 −309 A N ATOM 150 CA TYR A 64 42.981 14.803 39.034 1.00 31.59 A CANISOU 150 CA TYR A 64 4008 5088 2906 973 −1342 −467 A C ATOM 151 C TYRA 64 42.871 13.361 38.587 1.00 30.73 A C ANISOU 151 C TYR A 64 3980 52582440 1031 −1207 −692 A C ATOM 152 O TYR A 64 41.800 12.882 38.238 1.0036.32 A O ANISOU 152 O TYR A 64 4739 6072 2988 1075 −1224 −850 A O ATOM153 CB TYR A 64 42.805 14.863 40.542 1.00 29.64 A C ANISOU 153 CB TYR A64 3839 4503 2920 944 −1384 −725 A C ATOM 154 CG TYR A 64 44.008 14.41441.308 1.00 28.55 A C ANISOU 154 CG TYR A 64 3724 4304 2819 898 −1291−793 A C ATOM 155 CD1 TYR A 64 44.259 13.067 41.527 1.00 27.03 A CANISOU 155 CD1 TYR A 64 3620 4244 2407 937 −1161 −1041 A C ATOM 156 CD2TYR A 64 44.907 15.346 41.830 1.00 29.18 A C ANISOU 156 CD2 TYR A 643736 4188 3161 814 −1339 −603 A C ATOM 157 CE1 TYR A 64 45.391 12.66642.225 1.00 26.20 A C ANISOU 157 CE1 TYR A 64 3523 4091 2341 905 −1079−1082 A C ATOM 158 CE2 TYR A 64 46.007 14.952 42.521 1.00 28.32 A CANISOU 158 CE2 TYR A 64 3633 4046 3081 768 −1260 −652 A C ATOM 159 CZTYR A 64 46.254 13.613 42.720 1.00 26.82 A C ANISOU 159 CZ TYR A 64 35204002 2670 820 −1130 −886 A C ATOM 160 OH TYR A 64 47.381 13.238 43.4201.00 26.12 A O ANISOU 160 OH TYR A 64 3425 3884 2617 783 −1055 −914 A OATOM 161 N ARC A 65 43.984 12.662 38.620 1.00 30.38 A N ANISOU 161 N ARCA 65 3946 5323 2273 1031 −1075 −710 A N ATOM 162 CA ARC A 65 44.06811.316 38.084 1.00 33.86 A C ANISOU 162 CA ARC A 65 4446 5955 2464 1054−889 −855 A C ATOM 163 C ARC A 65 44.681 10.402 39.134 1.00 41.31 A CANISOU 163 C ARC A 65 5451 6680 3567 1006 −743 −1025 A C ATOM 164 O ARCA 65 45.781 10.668 39.626 1.00 51.64 A O ANISOU 164 O ARC A 65 6736 80074877 1031 −767 −989 A O ATOM 165 CB ARC A 65 44.889 11.309 36.788 1.0042.50 A C ANISOU 165 CB ARC A 65 5448 7410 3290 1089 −824 −606 A C ATOM166 CG ARC A 65 44.085 11.668 35.528 1.00 34.03 A C ANISOU 166 CG ARC A65 4317 6541 2072 1091 −869 −439 A C ATOM 167 CD ARC A 65 44.997 12.18334.411 1.00 40.56 A C ANISOU 167 CD ARC A 65 5004 7694 2713 1101 −856−77 A C ATOM 168 NE ARC A 65 44.226 12.503 33.222 1.00 52.64 A N ANISOU168 NE ARC A 65 6475 9397 4128 1085 −889 83 A N ATOM 169 CZ ARC A 6543.484 11.604 32.576 1.00 68.98 A C ANISOU 169 CZ ARC A 65 8607 115036098 1066 −780 −102 A C ATOM 170 NH1 ARC A 65 43.423 10.355 33.030 1.0066.13 A N1+ ANISOU 170 NH1 ARC A 65 8358 10972 5797 1048 −639 −406 A N1+ATOM 171 NH2 ARC A 65 42.794 11.941 31.489 1.00 79.93 A N ANISOU 171 NH2ARC A 65 9933 13063 7371 1049 −822 48 A N ATOM 172 N ALA A 66 43.9629.339 39.487 1.00 43.87 A N ANISOU 172 N ALA A 66 7435 3386 5848 −754216 −677 A N ATOM 173 CA ALA A 66 44.450 8.408 40.490 1.00 45.88 A CANISOU 173 CA ALA A 66 7716 3568 6149 −692 181 −669 A C ATOM 174 C ALA A66 45.790 7.821 40.058 1.00 53.76 A C ANISOU 174 C ALA A 66 8782 44557189 −527 228 −711 A C ATOM 175 O ALA A 66 45.982 7.460 38.893 1.0059.13 A O ANISOU 175 O ALA A 66 9596 5089 7780 −507 285 −769 A O ATOM176 CB ALA A 66 43.419 7.300 40.716 1.00 40.04 A C ANISOU 176 CB ALA A66 7103 2812 5296 −851 128 −672 A C ATOM 177 N GLN A 67 46.727 7.81841.001 1.00 47.91 A N ANISOU 177 N GLN A 67 7951 3671 6580 −404 204 −680A N ATOM 178 CA GLN A 67 48.086 7.348 40.794 1.00 44.34 A C ANISOU 178CA GLN A 67 7515 3123 6210 −232 245 −700 A C ATOM 179 C GLN A 67 48.2435.998 41.449 1.00 47.46 A C ANISOU 179 C GLN A 67 8003 3422 6607 −216207 −720 A C ATOM 180 O GLN A 67 47.653 5.733 42.485 1.00 39.50 A OANISOU 180 O GLN A 67 6979 2427 5604 −294 142 −686 A O ATOM 181 CB GLN A67 49.094 8.336 41.366 1.00 42.20 A C ANISOU 181 CB GLN A 67 7045 28666123 −99 227 −639 A C ATOM 182 CG GLN A 67 49.000 9.737 40.786 1.0050.85 A C ANISOU 182 CG GLN A 67 8037 4032 7251 −84 266 −605 A C ATOM183 CD GLN A 67 49.956 10.708 41.451 1.00 62.56 A C ANISOU 183 CD GLN A67 9331 5516 8923 40 195 −528 A C ATOM 184 OE1 GLN A 67 51.122 10.39241.671 1.00 75.67 A O ANISOU 184 OE1 GLN A 67 10969 7105 10678 181 192−508 A O ATOM 185 NE2 GLN A 67 49.466 11.894 41.773 1.00 61.83 A NANISOU 185 NE2 GLN A 67 9119 5501 8872 −10 118 −484 A N ATOM 186 N SER A68 49.053 5.149 40.837 1.00 58.67 A N ANISOU 186 N SER A 68 9521 47408029 −95 260 −768 A N ATOM 187 CA SER A 68 49.216 3.783 41.301 1.0067.34 A C ANISOU 187 CA SER A 68 10770 5724 9091 −86 238 −810 A C ATOM188 C SER A 68 49.768 3.559 42.702 1.00 61.51 A C ANISOU 188 C SER A 689951 4947 8473 −67 167 −760 A C ATOM 189 O SER A 68 49.242 2.730 43.4361.00 71.08 A O ANISOU 189 O SER A 68 11287 6096 9624 −145 124 −771 A OATOM 190 CB SER A 68 50.118 3.031 40.314 1.00 78.31 A C ANISOU 190 CBSER A 68 12272 7013 10471 78 326 −874 A C ATOM 191 OG SER A 68 49.9573.503 38.985 1.00 76.80 A O ANISOU 191 OG SER A 68 12219 6837 10123 59405 −934 A O ATOM 192 N PRO A 69 50.846 4.252 43.073 1.00 60.66 A NANISOU 192 N PRO A 69 9651 4861 8538 31 146 −703 A N ATOM 193 CA PRO A69 51.344 3.980 44.427 1.00 52.69 A C ANISOU 193 CA PRO A 69 8586 38137619 31 83 −658 A C ATOM 194 C PRO A 69 50.578 4.510 45.650 1.00 51.56 AC ANISOU 194 C PRO A 69 8364 3770 7455 −79 37 −601 A C ATOM 195 O PRO A69 50.237 3.724 46.522 1.00 44.25 A O ANISOU 195 O PRO A 69 7544 28396429 −190 8 −588 A O ATOM 196 CB PRO A 69 52.747 4.602 44.414 1.00 53.75A C ANISOU 196 CB PRO A 69 8572 3895 7955 199 85 −630 A C ATOM 197 CGPRO A 69 53.025 4.983 42.997 1.00 54.85 A C ANISOU 197 CG PRO A 69 86144104 8122 255 112 −622 A C ATOM 198 CD PRO A 69 51.701 5.188 42.339 1.0053.14 A C ANISOU 198 CD PRO A 69 8557 3927 7704 168 180 −680 A C ATOM199 N ASN A 70 50.243 5.796 45.681 1.00 47.40 A N ANISOU 199 N ASN A 707668 3328 7013 −47 32 −566 A N ATOM 200 CA ASN A 70 49.609 6.362 46.8721.00 49.75 A C ANISOU 200 CA ASN A 70 7881 3701 7321 −98 −3 −516 A CATOM 201 C ASN A 70 48.268 7.030 46.712 1.00 53.44 A C ANISOU 201 C ASNA 70 8381 4272 7652 −228 −5 −513 A C ATOM 202 O ASN A 70 47.674 7.44147.696 1.00 56.95 A O ANISOU 202 O ASN A 70 8798 4768 8073 −269 −30 −477A O ATOM 203 CB ASN A 70 50.524 7.421 47.486 1.00 53.92 A C ANISOU 203CB ASN A 70 8208 4249 8029 6 −1 −478 A C ATOM 204 CG ASN A 70 51.9857.063 47.395 1.00 55.37 A C ANISOU 204 CG ASN A 70 8333 4348 8359 128 18−460 A C ATOM 205 OD1 ASN A 70 52.468 6.200 48.114 1.00 56.47 A O ANISOU205 OD1 ASN A 70 8371 4503 8583 220 23 −446 A O ATOM 206 ND2 ASN A 7052.703 7.748 46.521 1.00 51.47 A N ANISOU 206 ND2 ASN A 70 7917 37667875 132 15 −451 A N ATOM 207 N PHE A 71 47.784 7.148 45.491 1.00 58.34A N ANISOU 207 N PHE A 71 9064 4922 8181 −289 28 −548 A N ATOM 208 CAPHE A 71 46.544 7.868 45.272 1.00 43.36 A C ANISOU 208 CA PHE A 71 71513129 6195 −403 32 −536 A C ATOM 209 C PHE A 71 45.328 7.092 44.798 1.0041.88 A C ANISOU 209 C PHE A 71 7120 2944 5848 −559 47 −559 A C ATOM 210O PHE A 71 44.518 7.608 44.047 1.00 45.19 A O ANISOU 210 O PHE A 71 75463425 6200 −639 71 −574 A O ATOM 211 CB PHE A 71 46.809 9.130 44.466 1.0034.21 A C ANISOU 211 CB PHE A 71 5866 2022 5109 −340 51 −538 A C ATOM212 CG PHE A 71 47.746 10.073 45.156 1.00 45.20 A C ANISOU 212 CG PHE A71 7083 3419 6672 −221 12 −500 A C ATOM 213 CD2 PHE A 71 49.109 9.89045.083 1.00 44.16 A C ANISOU 213 CD2 PHE A 71 6874 3215 6690 −86 4 −493A C ATOM 214 CD1 PHE A 71 47.262 11.114 45.916 1.00 53.96 A C ANISOU 214CD1 PHE A 71 8111 4595 7796 −247 −14 −472 A C ATOM 215 CE2 PHE A 7149.972 10.745 45.730 1.00 37.67 A C ANISOU 215 CE2 PHE A 71 5874 24356005 −1 −10 −438 A C ATOM 216 CE1 PHE A 71 48.121 11.975 46.567 1.0039.41 A C ANISOU 216 CE1 PHE A 71 6117 2756 6100 −147 −18 −441 A C ATOM217 CZ PHE A 71 49.478 11.792 46.473 1.00 28.09 A C ANISOU 217 CZ PHE A71 4595 1302 4774 −39 17 −413 A C ATOM 218 N LEU A 72 45.232 5.83645.207 1.00 39.64 A N ANISOU 218 N LEU A 72 6964 2591 5508 −617 23 −553A N ATOM 219 CA LEU A 72 44.085 5.014 44.845 1.00 43.24 A C ANISOU 219CA LEU A 72 7562 3038 5828 −789 9 −555 A C ATOM 220 C LEU A 72 42.9045.133 45.806 1.00 51.37 A C ANISOU 220 C LEU A 72 8597 4125 6795 −935−14 −486 A C ATOM 221 O LEU A 72 41.952 4.360 45.674 1.00 66.20 A OANISOU 221 O LEU A 72 10584 5988 8581 −1094 −38 −466 A O ATOM 222 CB LEUA 72 44.480 3.539 44.769 1.00 44.76 A C ANISOU 222 CB LEU A 72 7911 31005997 −781 −14 −583 A C ATOM 223 CG LEU A 72 45.483 3.108 43.701 1.0042.58 A C ANISOU 223 CG LEU A 72 7693 2741 5746 −660 17 −659 A C ATOM224 CD1 LEU A 72 45.339 3.996 42.513 1.00 42.61 A C ANISOU 224 CD1 LEU A72 7659 2816 5716 −656 64 −695 A C ATOM 225 CD2 LEU A 72 46.868 3.18944.234 1.00 53.14 A C ANISOU 225 CD2 LEU A 72 8941 4026 7224 −480 30−656 A C ATOM 226 N SER A 73 42.930 6.042 46.782 1.00 46.27 A N ANISOU226 N SER A 73 7851 3536 6194 −889 −10 −445 A N ATOM 227 CA SER A 7341.781 6.273 47.641 1.00 32.79 A C ANISOU 227 CA SER A 73 6164 1882 4411−1025 −12 −380 A C ATOM 228 C SER A 73 41.442 7.750 47.627 1.00 33.18 AC ANISOU 228 C SER A 73 6083 2038 4487 −1009 13 −375 A C ATOM 229 O SERA 73 42.331 8.610 47.605 1.00 38.74 A O ANISOU 229 O SER A 73 6677 27525289 −855 11 −403 A O ATOM 230 CB SER A 73 42.029 5.826 49.083 1.0046.84 A C ANISOU 230 CB SER A 73 8013 3601 6182 −996 −34 −330 A C ATOM231 OG SER A 73 42.698 6.829 49.822 1.00 44.92 A O ANISOU 231 OG SER A73 7676 3381 6010 −853 −38 −332 A O ATOM 232 N LEU A 74 40.142 8.03947.658 1.00 31.62 A N ANISOU 232 N LEU A 74 5888 1915 4210 −1178 32 −333A N ATOM 233 CA LEU A 74 39.715 9.428 47.794 1.00 35.61 A C ANISOU 233CA LEU A 74 6284 2512 4736 −1174 58 −325 A C ATOM 234 C LEU A 74 40.22410.065 49.090 1.00 42.75 A C ANISOU 234 C LEU A 74 7185 3385 5672 −104451 −314 A C ATOM 235 O LEU A 74 40.401 11.288 49.154 1.00 46.89 A OANISOU 235 O LEU A 74 7613 3947 6255 −954 49 −336 A O ATOM 236 CB LEU A74 38.196 9.519 47.730 1.00 40.21 A C ANISOU 236 CB LEU A 74 6860 31835234 −1399 87 −265 A C ATOM 237 CG LEU A 74 37.577 9.108 46.405 1.0048.99 A C ANISOU 237 CG LEU A 74 7952 4346 6317 −1533 69 −274 A C ATOM238 CD1 LEU A 74 36.079 9.229 46.463 1.00 50.42 A C ANISOU 238 CD1 LEU A74 8075 4640 6441 −1761 85 −190 A C ATOM 239 CD2 LEU A 74 38.126 9.99445.296 1.00 60.72 A C ANISOU 239 CD2 LEU A 74 9341 5867 7862 −1430 71−341 A C ATOM 240 N SER A 75 40.437 9.276 50.151 1.00 44.10 A N ANISOU240 N SER A 75 7473 3480 5801 −1033 36 −280 A N ATOM 241 CA SER A 7540.966 9.877 51.376 1.00 41.91 A C ANISOU 241 CA SER A 75 7217 3169 5539−899 12 −275 A C ATOM 242 C SER A 75 42.369 10.426 51.139 1.00 44.10 A CANISOU 242 C SER A 75 7359 3431 5964 −692 −38 −329 A C ATOM 243 O SER A75 42.684 11.551 51.550 1.00 37.81 A O ANISOU 243 O SER A 75 6487 26565224 −581 −66 −346 A O ATOM 244 CB SER A 75 40.943 8.876 52.533 1.0046.11 A C ANISOU 244 CB SER A 75 7916 3618 5984 −935 1 −218 A C ATOM 245OG SER A 75 41.528 7.639 52.175 1.00 58.18 A O ANISOU 245 OG SER A 759479 5083 7542 −933 −27 −225 A O ATOM 246 N ASN A 76 43.189 9.688 50.3771.00 47.35 A N ANISOU 246 N ASN A 76 7735 3808 6448 −649 −46 −355 A NATOM 247 CA ASN A 76 44.579 10.085 50.134 1.00 41.60 A C ANISOU 247 CAASN A 76 6872 3057 5876 −482 −71 −387 A C ATOM 248 C ASN A 76 44.68311.368 49.298 1.00 33.03 A C ANISOU 248 C ASN A 76 5633 2037 4880 −434−62 −411 A C ATOM 249 O ASN A 76 45.577 12.195 49.541 1.00 30.27 A OANISOU 249 O ASN A 76 5160 1685 4656 −312 −78 −415 A O ATOM 250 CB ASN A76 45.338 8.922 49.492 1.00 40.88 A C ANISOU 250 CB ASN A 76 6812 28975826 −461 −63 −406 A C ATOM 251 CG ASN A 76 45.993 8.026 50.530 1.0055.01 A C ANISOU 251 CG ASN A 76 8677 4600 7623 −414 −93 −383 A C ATOM252 OD1 ASN A 76 45.516 7.922 51.650 1.00 60.05 A O ANISOU 252 OD1 ASN A76 9411 5229 8174 −451 −117 −345 A O ATOM 253 ND2 ASN A 76 47.101 7.39050.164 1.00 68.33 A N ANISOU 253 ND2 ASN A 76 10337 6217 9410 −331 −90−403 A N ATOM 254 N ILE A 77 43.795 11.546 48.305 1.00 30.11 A N ANISOU254 N ILE A 77 5270 1723 4449 −542 −29 −421 A N ATOM 255 CA ILE A 7743.771 12.767 47.488 1.00 26.61 A C ANISOU 255 CA ILE A 77 4700 13374076 −514 −25 −435 A C ATOM 256 C ILE A 77 43.168 13.923 48.267 1.0028.36 A C ANISOU 256 C ILE A 77 4885 1600 4291 −505 −44 −424 A C ATOM257 O ILE A 77 43.596 15.095 48.173 1.00 26.44 A O ANISOU 257 O ILE A 774517 1371 4157 −411 −67 −432 A O ATOM 258 CB ILE A 77 42.975 12.50446.201 1.00 35.47 A C ANISOU 258 CB ILE A 77 5862 2503 5113 −648 18 −446A C ATOM 259 CG1 ILE A 77 43.558 11.298 45.464 1.00 27.74 A C ANISOU 259CG1 ILE A 77 4966 1461 4114 −642 35 −472 A C ATOM 260 CG2 ILE A 7742.886 13.769 45.354 1.00 26.02 A C ANISOU 260 CG2 ILE A 77 4549 13623974 −634 23 −451 A C ATOM 261 CD1 ILE A 77 42.601 10.698 44.478 1.0028.51 A C ANISOU 261 CD1 ILE A 77 5159 1588 4087 −801 65 −487 A C ATOM262 N SER A 78 42.124 13.603 49.009 1.00 35.73 A N ANISOU 262 N SER A 785939 2544 5093 −613 −26 −404 A N ATOM 263 CA SER A 78 41.533 14.53049.955 1.00 40.34 A C ANISOU 263 CA SER A 78 6551 3139 5638 −595 −37−400 A C ATOM 264 C SER A 78 42.587 15.111 50.903 1.00 35.58 A C ANISOU264 C SER A 78 5904 2493 5123 −404 −110 −413 A C ATOM 265 O SER A 7842.535 16.297 51.274 1.00 27.11 A O ANISOU 265 O SER A 78 4783 1433 4084−316 −151 −431 A O ATOM 266 CB SER A 78 40.454 13.779 50.726 1.00 47.62A C ANISOU 266 CB SER A 78 7645 4050 6400 −748 17 −359 A C ATOM 267 OGSER A 78 39.876 14.626 51.695 1.00 66.19 A O ANISOU 267 OG SER A 7810100 6354 8698 −661 −5 −356 A O ATOM 268 N ASP A 79 43.571 14.29151.285 1.00 31.96 A N ANISOU 268 N ASP A 79 5457 1985 4703 −344 −128−404 A N ATOM 269 CA ASP A 79 44.591 14.738 52.231 1.00 37.06 A C ANISOU269 CA ASP A 79 6067 2602 5413 −200 −181 −403 A C ATOM 270 C ASP A 7945.445 15.866 51.663 1.00 44.41 A C ANISOU 270 C ASP A 79 6832 3565 6475−109 −173 −405 A C ATOM 271 O ASP A 79 45.833 16.781 52.403 1.00 51.82 AO ANISOU 271 O ASP A 79 7757 4516 7418 −14 −236 −397 A O ATOM 272 CB ASPA 79 45.467 13.559 52.668 1.00 42.08 A C ANISOU 272 CB ASP A 79 67533175 6060 −180 −187 −382 A C ATOM 273 CG ASP A 79 44.736 12.602 53.6241.00 61.16 A C ANISOU 273 CG ASP A 79 9378 5547 8314 −253 −201 −356 A CATOM 274 OD1 ASP A 79 43.692 12.988 54.198 1.00 65.98 A O ANISOU 274 OD1ASP A 79 10106 6167 8797 −298 −198 −349 A O ATOM 275 OD2 ASP A 79 45.20811.465 53.841 1.00 65.70 A O1− ANISOU 275 OD2 ASP A 79 10017 6064 8880−268 −204 −336 A O1− ATOM 276 N ILE A 80 45.744 15.842 50.360 1.00 39.50A N ANISOU 276 N ILE A 80 6122 2958 5930 −139 −103 −405 A N ATOM 277 CAILE A 80 46.558 16.927 49.814 1.00 34.04 A C ANISOU 277 CA ILE A 80 53582296 5280 −63 −87 −372 A C ATOM 278 C ILE A 80 45.721 18.093 49.307 1.0028.37 A C ANISOU 278 C ILE A 80 4593 1640 4548 −81 −124 −379 A C ATOM279 O ILE A 80 46.294 19.135 48.960 1.00 23.66 A O ANISOU 279 O ILE A 803910 1082 3996 −9 −189 −342 A O ATOM 280 CB ILE A 80 47.508 16.42648.708 1.00 35.19 A C ANISOU 280 CB ILE A 80 5498 2411 5460 −54 −3 −348A C ATOM 281 CG1 ILE A 80 46.761 16.162 47.409 1.00 41.25 A C ANISOU 281CG1 ILE A 80 6235 3218 6221 −148 51 −375 A C ATOM 282 CG2 ILE A 8048.172 15.132 49.138 1.00 33.60 A C ANISOU 282 CG2 ILE A 80 5319 21505299 −42 25 −351 A C ATOM 283 CD1 ILE A 80 47.638 15.507 46.337 1.0026.52 A C ANISOU 283 CD1 ILE A 80 4384 1326 4366 −125 124 −360 A C ATOM284 N PHE A 81 44.386 17.970 49.258 1.00 33.24 A N ANISOU 284 N PHE A 815235 2267 5128 −175 −120 −418 A N ATOM 285 CA PHE A 81 43.590 19.12048.829 1.00 32.09 A C ANISOU 285 CA PHE A 81 5033 2171 4988 −186 −167−423 A C ATOM 286 C PHE A 81 42.785 19.835 49.907 1.00 46.01 A C ANISOU286 C PHE A 81 6827 3935 6719 −132 −250 −452 A C ATOM 287 O PHE A 8142.121 20.821 49.574 1.00 52.42 A O ANISOU 287 O PHE A 81 7557 4782 7579−120 −302 −466 A O ATOM 288 CB PHE A 81 42.640 18.727 47.706 1.00 26.08A C ANISOU 288 CB PHE A 81 4269 1427 4213 −342 −118 −437 A C ATOM 289 CGPHE A 81 43.339 18.484 46.396 1.00 23.48 A C ANISOU 289 CG PHE A 81 39151100 3906 −364 −60 −416 A C ATOM 290 CD1 PHE A 81 43.894 19.545 45.6751.00 22.98 A C ANISOU 290 CD1 PHE A 81 3724 1081 3926 −287 −119 −374 A CATOM 291 CD2 PHE A 81 43.419 17.209 45.872 1.00 23.98 A C ANISOU 291 CD2PHE A 81 3983 1138 3990 −421 −63 −430 A C ATOM 292 CE1 PHE A 81 44.52419.333 44.513 1.00 23.01 A C ANISOU 292 CE1 PHE A 81 3707 1077 3960 −292−84 −349 A C ATOM 293 CE2 PHE A 81 44.049 16.997 44.682 1.00 30.82 A CANISOU 293 CE2 PHE A 81 4844 1996 4872 −409 −72 −413 A C ATOM 294 CZ PHEA 81 44.614 18.061 44.006 1.00 25.71 A C ANISOU 294 CZ PHE A 81 41981361 4210 −360 −45 −370 A C ATOM 295 N ASN A 82 42.785 19.367 51.1611.00 44.06 A N ANISOU 295 N ASN A 82 6698 3646 6395 −90 −271 −466 A NATOM 296 CA ASN A 82 42.104 20.067 52.266 1.00 43.67 A C ANISOU 296 CAASN A 82 6735 3585 6274 1 −345 −502 A C ATOM 297 C ASN A 82 40.58820.057 52.119 1.00 38.27 A C ANISOU 297 C ASN A 82 6035 3043 5463 −95−246 −492 A C ATOM 298 O ASN A 82 39.937 21.107 52.174 1.00 45.53 A OANISOU 298 O ASN A 82 6839 4082 6377 −22 −251 −503 A O ATOM 299 CB ASN A82 42.558 21.522 52.413 1.00 52.07 A C ANISOU 299 CB ASN A 82 7669 46917424 146 −442 −510 A C ATOM 300 CG ASN A 82 43.950 21.665 52.949 1.0050.89 A C ANISOU 300 CG ASN A 82 7483 4544 7308 223 −500 −472 A C ATOM301 OD1 ASN A 82 44.602 20.685 53.340 1.00 53.99 A O ANISOU 301 OD1 ASNA 82 7939 4899 7675 199 −470 −451 A O ATOM 302 ND2 ASN A 82 44.41522.912 52.990 1.00 42.82 A N ANISOU 302 ND2 ASN A 82 6358 3561 6351 311−602 −461 A N ATOM 303 N LEU A 83 40.021 18.875 51.909 1.00 27.33 A NANISOU 303 N LEU A 83 4706 1705 3973 −261 −134 −450 A N ATOM 304 CA LEUA 83 38.567 18.731 51.894 1.00 28.54 A C ANISOU 304 CA LEU A 83 47832064 3996 −372 −12 −399 A C ATOM 305 C LEU A 83 38.237 17.257 52.1011.00 30.85 A C ANISOU 305 C LEU A 83 5210 2335 4178 −539 67 −345 A CATOM 306 O LEU A 83 39.120 16.402 52.113 1.00 31.30 A O ANISOU 306 O LEUA 83 5412 2216 4263 −562 28 −360 A O ATOM 307 CB LEU A 83 37.942 19.29450.601 1.00 41.10 A C ANISOU 307 CB LEU A 83 6172 3793 5653 −456 17 −381A C ATOM 308 CG LEU A 83 38.271 18.721 49.210 1.00 33.52 A C ANISOU 308CG LEU A 83 5162 2756 4820 −514 −21 −396 A C ATOM 309 CD1 LEU A 8337.498 17.431 48.874 1.00 29.44 A C ANISOU 309 CD1 LEU A 83 4772 21544261 −675 24 −384 A C ATOM 310 CD2 LEU A 83 38.041 19.792 48.152 1.0026.22 A C ANISOU 310 CD2 LEU A 83 4034 1992 3938 −554 −8 −372 A C ATOM311 N SER A 84 36.962 16.965 52.283 1.00 31.49 A N ANISOU 311 N SER A 845234 2589 4143 −653 173 −273 A N ATOM 312 CA SER A 84 36.576 15.60452.613 1.00 32.35 A C ANISOU 312 CA SER A 84 5468 2675 4149 −819 237−201 A C ATOM 313 C SER A 84 36.619 14.713 51.373 1.00 29.45 A C ANISOU313 C SER A 84 5112 2243 3833 −1015 229 −194 A C ATOM 314 O SER A 8436.458 15.192 50.252 1.00 40.82 A O ANISOU 314 O SER A 84 6418 3744 5346−1055 216 −216 A O ATOM 315 CB SER A 84 35.177 15.616 53.224 1.00 43.32A C ANISOU 315 CB SER A 84 6768 4282 5410 −880 356 −101 A C ATOM 316 OGSER A 84 34.304 16.553 52.566 1.00 49.11 A O ANISOU 316 OG SER A 84 72695208 6184 −881 391 −86 A O ATOM 317 N PRO A 85 36.809 13.403 51.535 1.0030.18 A N ANISOU 317 N PRO A 85 5378 2209 3881 −1139 231 −165 A N ATOM318 CA PRO A 85 36.553 12.512 50.394 1.00 30.48 A C ANISOU 318 CA PRO A85 5440 2206 3936 −1343 226 −153 A C ATOM 319 C PRO A 85 35.150 12.66249.838 1.00 50.41 A C ANISOU 319 C PRO A 85 7789 4939 6425 −1513 276 −74A C ATOM 320 O PRO A 85 34.921 12.361 48.662 1.00 58.33 A O ANISOU 320 OPRO A 85 8766 5939 7458 −1651 246 −86 A O ATOM 321 CB PRO A 85 36.76311.104 50.969 1.00 31.60 A C ANISOU 321 CB PRO A 85 5804 2187 4017 −1446219 −114 A C ATOM 322 CG PRO A 85 37.378 11.264 52.291 1.00 31.62 A CANISOU 322 CG PRO A 85 5906 2123 3986 −1278 209 −116 A C ATOM 323 CD PROA 85 37.086 12.657 52.774 1.00 34.36 A C ANISOU 323 CD PRO A 85 60932637 4325 −1113 235 −128 A C ATOM 324 N LEU A 86 34.187 13.100 50.6521.00 51.28 A N ANISOU 324 N LEU A 86 7779 5234 6470 −1502 351 12 A NATOM 325 CA LEU A 86 32.821 13.248 50.160 1.00 54.62 A C ANISOU 325 CALEU A 86 8006 5867 6879 −1660 399 108 A C ATOM 326 C LEU A 86 32.76814.251 49.024 1.00 44.39 A C ANISOU 326 C LEU A 86 6543 4653 5672 −1619358 49 A C ATOM 327 O LEU A 86 32.116 14.015 48.004 1.00 39.57 A OANISOU 327 O LEU A 86 5849 4105 5081 −1795 331 84 A O ATOM 328 CB LEU A86 31.882 13.693 51.274 1.00 59.57 A C ANISOU 328 CB LEU A 86 8513 66947426 −1601 509 210 A C ATOM 329 CG LEU A 86 30.438 13.975 50.857 1.0060.40 A C ANISOU 329 CG LEU A 86 8370 7043 7538 −1736 569 328 A C ATOM330 CD1 LEU A 86 29.766 12.699 50.354 1.00 48.56 A C ANISOU 330 CD1 LEUA 86 6895 5522 6035 −2043 546 439 A C ATOM 331 CD2 LEU A 86 29.69814.543 52.037 1.00 77.58 A C ANISOU 331 CD2 LEU A 86 10434 9413 9630−1607 700 414 A C ATOM 332 N ARC A 87 33.420 15.397 49.197 1.00 30.63 AN ANISOU 332 N ARC A 87 4749 2907 3980 −1392 340 −32 A N ATOM 333 CA ARCA 87 33.286 16.438 48.198 1.00 29.78 A C ANISOU 333 CA ARC A 87 44672891 3956 −1349 304 −65 A C ATOM 334 C ARC A 87 33.850 15.984 46.8651.00 45.48 A C ANISOU 334 C ARC A 87 6521 4765 5994 −1452 236 −116 A CATOM 335 O ARC A 87 33.266 16.254 45.805 1.00 49.67 A O ANISOU 335 O ARCA 87 6929 5396 6547 −1554 213 −95 A O ATOM 336 CB ARC A 87 33.975 17.70048.667 1.00 28.72 A C ANISOU 336 CB ARC A 87 4291 2739 3882 −1093 274−138 A C ATOM 337 CG ARC A 87 33.175 18.461 49.634 1.00 29.48 A C ANISOU337 CG ARC A 87 4278 2997 3927 −976 338 −98 A C ATOM 338 CD ARC A 8733.829 19.788 49.921 1.00 41.62 A C ANISOU 338 CD ARC A 87 5781 44975535 −730 273 −182 A C ATOM 339 NE ARC A 87 33.793 20.718 48.796 1.0043.59 A N ANISOU 339 NE ARC A 87 5867 4797 5899 −717 215 −199 A N ATOM340 CZ ARC A 87 34.089 22.012 48.925 1.00 48.77 A C ANISOU 340 CZ ARC A87 6447 5450 6633 −525 152 −249 A C ATOM 341 NH1 ARC A 87 34.031 22.81247.864 1.00 41.79 A N1+ ANISOU 341 NH1 ARC A 87 5415 4610 5854 −530 97−247 A N1+ ATOM 342 NH2 ARC A 87 34.435 22.507 50.132 1.00 48.53 A NANISOU 342 NH2 ARC A 87 6504 5365 6570 −328 131 −299 A N ATOM 343 N ILEA 88 34.978 15.273 46.903 1.00 42.69 A N ANISOU 343 N ILE A 88 6367 42045649 −1420 206 −181 A N ATOM 344 CA ILE A 88 35.557 14.744 45.678 1.0037.36 A C ANISOU 344 CA ILE A 88 5782 3415 4998 −1494 163 −234 A C ATOM345 C ILE A 88 34.618 13.710 45.067 1.00 41.23 A C ANISOU 345 C ILE A 886318 3932 5414 −1748 153 −183 A C ATOM 346 O ILE A 88 34.350 13.73743.862 1.00 46.70 A O ANISOU 346 O ILE A 88 6977 4662 6106 −1845 115−196 A O ATOM 347 CB ILE A 88 36.963 14.181 45.954 1.00 32.76 A C ANISOU347 CB ILE A 88 5394 2605 4448 −1381 144 −306 A C ATOM 348 CG1 ILE A 8837.858 15.293 46.479 1.00 26.83 A C ANISOU 348 CG1 ILE A 88 4573 18283792 −1147 122 −343 A C ATOM 349 CG2 ILE A 88 37.573 13.616 44.684 1.0037.72 A C ANISOU 349 CG2 ILE A 88 6073 3174 5085 −1379 116 −352 A C ATOM350 CD1 ILE A 88 39.257 14.821 46.815 1.00 31.57 A C ANISOU 350 CD1 ILEA 88 5304 2242 4450 −1008 88 −389 A C ATOM 351 N ALA A 89 34.044 12.82945.897 1.00 37.95 A N ANISOU 351 N ALA A 89 5978 3506 4936 −1868 176−113 A N ATOM 352 CA ALA A 89 33.108 11.828 45.379 1.00 32.60 A C ANISOU352 CA ALA A 89 5329 2850 4206 −2115 141 −46 A C ATOM 353 C ALA A 8931.901 12.475 44.709 1.00 37.73 A C ANISOU 353 C ALA A 89 5758 3710 4869−2245 130 27 A C ATOM 354 O ALA A 89 31.416 11.985 43.684 1.00 46.97 A OANISOU 354 O ALA A 89 6916 4908 6022 −2352 53 36 A O ATOM 355 CB ALA A89 32.644 10.881 46.486 1.00 33.98 A C ANISOU 355 CB ALA A 89 5586 29984326 −2219 172 50 A C ATOM 356 N LYS A 90 31.386 13.565 45.273 1.0036.57 A N ANISOU 356 N LYS A 90 5394 3750 4750 −2132 191 78 A N ATOM 357CA LYS A 90 30.193 14.172 44.695 1.00 37.29 A C ANISOU 357 CA LYS A 905249 4055 4866 −2231 179 161 A C ATOM 358 C LYS A 90 30.526 14.97543.450 1.00 44.55 A C ANISOU 358 C LYS A 90 6106 4991 5832 −2175 115 87A C ATOM 359 O LYS A 90 29.705 15.045 42.523 1.00 53.45 A O ANISOU 359 OLYS A 90 7124 6220 6964 −2326 53 137 A O ATOM 360 CB LYS A 90 29.46815.048 45.721 1.00 33.76 A C ANISOU 360 CB LYS A 90 4592 3806 4428 −2113276 244 A C ATOM 361 CG LYS A 90 28.806 14.247 46.864 1.00 49.80 A CANISOU 361 CG LYS A 90 6640 5884 6398 −2207 358 366 A C ATOM 362 CD LYSA 90 27.687 15.019 47.584 1.00 64.87 A C ANISOU 362 CD LYS A 90 82948049 8305 −2143 466 485 A C ATOM 363 CE LYS A 90 28.188 16.259 48.3421.00 73.33 A C ANISOU 363 CE LYS A 90 9327 9163 9373 −1829 529 404 A CATOM 364 NZ LYS A 90 27.077 16.921 49.102 1.00 76.22 A N1+ ANISOU 364 NZLYS A 90 9470 9774 9716 −1745 651 515 A N1+ ATOM 365 N ALA A 91 31.73015.550 43.385 1.00 37.90 A N ANISOU 365 N ALA A 91 5332 4044 5025 −1970118 −19 A N ATOM 366 CA ALA A 91 32.117 16.278 42.182 1.00 29.97 A CANISOU 366 CA ALA A 91 4275 3051 4062 −1918 66 −71 A C ATOM 367 C ALA A91 32.497 15.337 41.037 1.00 38.82 A C ANISOU 367 C ALA A 91 5584 40425125 −2051 6 −125 A C ATOM 368 O ALA A 91 32.286 15.662 39.856 1.0035.13 A O ANISOU 368 O ALA A 91 5070 3631 4647 −2106 −49 −130 A O ATOM369 CB ALA A 91 33.255 17.233 42.501 1.00 28.41 A C ANISOU 369 CB ALA A91 4067 2790 3939 −1666 86 −140 A C ATOM 370 N SER A 92 33.034 14.16541.363 1.00 34.42 A N ANISOU 370 N SER A 92 5243 3316 4520 −2076 11 −165A N ATOM 371 CA SER A 92 33.494 13.192 40.378 1.00 31.48 A C ANISOU 371CA SER A 92 5041 2839 4082 −2075 −39 −226 A C ATOM 372 C SER A 92 32.39712.252 39.890 1.00 38.94 A C ANISOU 372 C SER A 92 6008 3830 4955 −2264−116 −172 A C ATOM 373 O SER A 92 32.632 11.476 38.957 1.00 45.25 A OANISOU 373 O SER A 92 6961 4548 5685 −2287 −172 −227 A O ATOM 374 CB SERA 92 34.639 12.369 40.978 1.00 40.72 A C ANISOU 374 CB SER A 92 63943829 5248 −1938 −6 −288 A C ATOM 375 OG SER A 92 35.686 13.225 41.4401.00 35.93 A O ANISOU 375 OG SER A 92 5758 3170 4723 −1751 46 −329 A OATOM 376 N ASN A 93 31.213 12.298 40.505 1.00 41.81 A N ANISOU 376 N ASNA 93 6226 4321 5339 −2399 −120 −61 A N ATOM 377 CA ASN A 93 30.10011.386 40.212 1.00 37.39 A C ANISOU 377 CA ASN A 93 5663 3804 4741 −2585−202 16 A C ATOM 378 C ASN A 93 30.435 9.944 40.609 1.00 42.72 A CANISOU 378 C ASN A 93 6550 4312 5368 −2610 −223 −3 A C ATOM 379 O ASN A93 30.073 8.983 39.926 1.00 41.63 A O ANISOU 379 O ASN A 93 6522 41175180 −2717 −319 −6 A O ATOM 380 CB ASN A 93 29.661 11.484 38.748 1.0036.98 A C ANISOU 380 CB ASN A 93 5600 3802 4649 −2659 −305 −1 A C ATOM381 CG ASN A 93 28.337 10.795 38.481 1.00 48.90 A C ANISOU 381 CG ASN A93 7052 5382 6147 −2857 −406 100 A C ATOM 382 OD1 ASN A 93 27.369 10.90639.244 1.00 49.43 A O ANISOU 382 OD1 ASN A 93 6938 5571 6273 −2944 −383225 A O ATOM 383 ND2 ASN A 93 28.296 10.061 37.379 1.00 60.69 A N ANISOU383 ND2 ASN A 93 8705 6792 7561 −2930 −519 50 A N ATOM 384 N ILE A 9431.062 9.787 41.769 1.00 53.62 A N ANISOU 384 N ILE A 94 7995 5611 6766−2513 −143 −13 A N ATOM 385 CA ILE A 94 31.422 8.460 42.250 1.00 60.33 AC ANISOU 385 CA ILE A 94 9014 6322 7586 −2535 −154 −4 A C ATOM 386 C ILEA 94 30.409 7.959 43.276 1.00 62.25 A C ANISOU 386 C ILE A 94 9144 66607847 −2676 −119 145 A C ATOM 387 O ILE A 94 30.173 8.600 44.297 1.0059.20 A O ANISOU 387 O ILE A 94 8623 6379 7489 −2650 −26 202 A O ATOM388 CB ILE A 94 32.823 8.459 42.881 1.00 51.19 A C ANISOU 388 CB ILE A94 7994 5018 6437 −2327 −95 −98 A C ATOM 389 CG1 ILE A 94 33.858 8.92941.865 1.00 47.82 A C ANISOU 389 CG1 ILE A 94 7657 4515 5997 −2192 −114−220 A C ATOM 390 CG2 ILE A 94 33.176 7.075 43.391 1.00 51.23 A C ANISOU390 CG2 ILE A 94 8160 4888 6417 −2339 −109 −79 A C ATOM 391 CD1 ILE A 9435.281 8.828 42.357 1.00 50.65 A C ANISOU 391 CD1 ILE A 94 8079 47716395 −1977 −55 −290 A C ATOM 392 N GLU A 95 29.800 6.815 42.980 1.0064.94 A N ANISOU 392 N GLU A 95 9539 6966 8170 −2831 −189 218 A N ATOM393 CA GLU A 95 28.798 6.213 43.857 1.00 80.14 A C ANISOU 393 CA GLU A95 11343 8989 10116 −2973 −147 385 A C ATOM 394 C GLU A 95 29.319 5.76045.221 1.00 81.49 A C ANISOU 394 C GLU A 95 11677 9023 10261 −2960 −115413 A C ATOM 395 O GLU A 95 28.667 5.979 46.238 1.00 85.23 A O ANISOU395 O GLU A 95 12070 9574 10741 −3068 −60 565 A O ATOM 396 CB GLU A 9528.083 5.057 43.145 1.00 90.92 A C ANISOU 396 CB GLU A 95 12606 1043511504 −3177 −250 490 A C ATOM 397 CG GLU A 95 29.007 3.977 42.603 1.0099.86 A C ANISOU 397 CG GLU A 95 13949 11395 12600 −3245 −393 427 A CATOM 398 CD GLU A 95 28.267 2.922 41.804 1.00 107.46 A C ANISOU 398 CDGLU A 95 14812 12431 13587 −3446 −519 521 A C ATOM 399 OE1 GLU A 9528.832 1.828 41.595 1.00 116.17 A O ANISOU 399 OE1 GLU A 95 15891 1352714721 −3602 −558 652 A O ATOM 400 OE2 GLU A 95 27.121 3.188 41.384 1.00104.53 A O1− ANISOU 400 OE2 GLU A 95 14386 12124 13207 −3450 −584 473 AO1− ATOM 401 N ALA A 96 30.494 5.140 45.246 1.00 71.81 A N ANISOU 401 NALA A 96 10668 7610 9009 −2822 −142 283 A N ATOM 402 CA ALA A 96 31.0514.670 46.505 1.00 66.37 A C ANISOU 402 CA ALA A 96 10141 6778 8297 −2792−130 302 A C ATOM 403 C ALA A 96 32.259 5.498 46.890 1.00 73.06 A CANISOU 403 C ALA A 96 11011 7611 9137 −2635 −20 285 A C ATOM 404 O ALA A96 33.271 5.502 46.198 1.00 77.47 A O ANISOU 404 O ALA A 96 11602 81629669 −2679 35 389 A O ATOM 405 CB ALA A 96 31.432 3.205 46.396 1.0057.47 A C ANISOU 405 CB ALA A 96 9223 5461 7152 −2715 −220 181 A C ATOM406 N GLU A 97 32.150 6.179 48.020 1.00 65.72 A N ANISOU 406 N GLU A 9710071 6673 8227 −2454 7 163 A N ATOM 407 CA GLU A 97 33.216 7.029 48.5201.00 56.65 A C ANISOU 407 CA GLU A 97 8947 5493 7085 −2284 86 124 A CATOM 408 C GLU A 97 34.472 6.245 48.879 1.00 47.54 A C ANISOU 408 C GLUA 97 7976 4162 5925 −2141 60 69 A C ATOM 409 O GLU A 97 35.585 6.67848.606 1.00 40.15 A O ANISOU 409 O GLU A 97 7070 3156 5028 −1933 57 −37A O ATOM 410 CB GLU A 97 32.707 7.816 49.727 1.00 59.24 A C ANISOU 410CB GLU A 97 9191 5939 7380 −2374 190 256 A C ATOM 411 CG GLU A 97 33.6568.866 50.273 1.00 74.07 A C ANISOU 411 CG GLU A 97 11099 7784 9260 −2198263 197 A C ATOM 412 CD GLU A 97 32.990 9.742 51.316 1.00 93.12 A CANISOU 412 CD GLU A 97 13369 10401 11610 −2149 375 313 A C ATOM 413 OE1GLU A 97 33.630 10.058 52.340 1.00 90.17 A O ANISOU 413 OE1 GLU A 9713062 9999 11198 −1945 409 281 A O ATOM 414 OE2 GLU A 97 31.814 10.10551.116 1.00 106.91 A O1− ANISOU 414 OE2 GLU A 97 14933 12344 13343 −2296425 438 A O1− ATOM 415 N ASP A 98 34.280 5.082 49.489 1.00 53.16 A NANISOU 415 N ASP A 98 8787 4812 6599 −2250 38 154 A N ATOM 416 CA ASP A98 35.390 4.249 49.941 1.00 65.58 A C ANISOU 416 CA ASP A 98 10522 62258169 −2122 8 115 A C ATOM 417 C ASP A 98 35.971 3.263 48.934 1.00 61.88A C ANISOU 417 C ASP A 98 10134 5649 7730 −2056 −72 16 A C ATOM 418 OASP A 98 36.968 2.610 49.224 1.00 67.72 A O ANISOU 418 O ASP A 98 109686271 8490 −1905 −91 −38 A O ATOM 419 CB ASP A 98 34.987 3.496 51.2101.00 77.98 A C ANISOU 419 CB ASP A 98 12180 7762 9687 −2261 17 255 A CATOM 420 CG ASP A 98 34.719 4.424 52.374 1.00 93.26 A C ANISOU 420 CGASP A 98 14093 9774 11568 −2275 121 355 A C ATOM 421 OD1 ASP A 98 35.6515.141 52.792 1.00 89.20 A O ANISOU 421 OD1 ASP A 98 13623 9216 11053−2087 150 283 A O ATOM 422 OD2 ASP A 98 33.577 4.432 52.875 1.00 107.87A O1−− ANISOU 422 OD2 ASP A 98 15873 11736 13375 −2469 177 515 A O1 ATOM423 N LYS A 99 35.363 3.139 47.762 1.00 62.44 A N ANISOU 423 N LYS A 9910169 5757 7797 −2162 −118 −5 A N ATOM 424 CA LYS A 99 35.872 2.19346.776 1.00 64.48 A C ANISOU 424 CA LYS A 99 10545 5897 8056 −2122 −190−90 A C ATOM 425 C LYS A 99 37.267 2.572 46.296 1.00 57.96 A C ANISOU425 C LYS A 99 9725 5026 7273 −1878 −160 −214 A C ATOM 426 O LYS A 9937.553 3.743 46.071 1.00 56.91 A O ANISOU 426 O LYS A 99 9473 4980 7171−1780 −108 −248 A O ATOM 427 CB LYS A 99 34.916 2.081 45.589 1.00 69.66A C ANISOU 427 CB LYS A 99 11178 6604 8686 −2286 −253 −90 A C ATOM 428CG LYS A 99 35.383 1.111 44.516 1.00 77.75 A C ANISOU 428 CG LYS A 9912353 7501 9688 −2245 −326 −189 A C ATOM 429 CD LYS A 99 34.251 0.73243.578 1.00 80.79 A C ANISOU 429 CD LYS A 99 12765 7899 10033 −2455 −426−157 A C ATOM 430 CE LYS A 99 33.577 1.963 43.001 1.00 78.59 A C ANISOU430 CE LYS A 99 12333 7779 9747 −2514 −415 −154 A C ATOM 431 NZ LYS A 9932.376 1.606 42.199 1.00 84.63 A N1+ ANISOU 431 NZ LYS A 99 13114 855910481 −2730 −534 −107 A N1+ ATOM 432 N LYS A 100 38.133 1.577 46.1351.00 53.53 A N ANISOU 432 N LYS A 100 9290 4325 6722 −1782 −193 −269 A NATOM 433 CA LYS A 100 39.488 1.833 45.672 1.00 50.73 A C ANISOU 433 CALYS A 100 8925 3928 6422 −1558 −161 −368 A C ATOM 434 C LYS A 100 39.4712.305 44.219 1.00 58.40 A C ANISOU 434 C LYS A 100 9872 4945 7374 −1546−153 −444 A C ATOM 435 O LYS A 100 38.823 1.697 43.357 1.00 61.43 A OANISOU 435 O LYS A 100 10347 5301 7691 −1673 −208 −462 A O ATOM 436 CBLYS A 100 40.350 0.573 45.822 1.00 51.07 A C ANISOU 436 CB LYS A 1009109 3812 6483 −1476 −196 −402 A C ATOM 437 CG LYS A 100 41.860 0.82845.638 1.00 61.24 A C ANISOU 437 CG LYS A 100 10356 5060 7854 −1238 −153−478 A C ATOM 438 CD LYS A 100 42.737 −0.388 45.997 1.00 63.80 A CANISOU 438 CD LYS A 100 10800 5230 8213 −1154 −183 −497 A C ATOM 439 CELYS A 100 44.230 −0.064 45.824 1.00 68.69 A C ANISOU 439 CE LYS A 10011343 5821 8934 −927 −137 −558 A C ATOM 440 NZ LYS A 100 45.147 −1.18546.205 1.00 74.32 A N1+ ANISOU 440 NZ LYS A 100 12152 6387 9698 −836−162 −574 A N1+ ATOM 441 N LEU A 101 40.185 3.396 43.954 1.00 53.74 A NANISOU 441 N LEU A 101 9165 4416 6837 −1397 −94 −485 A N ATOM 442 CA LEUA 101 40.242 3.971 42.625 1.00 50.02 A C ANISOU 442 CA LEU A 101 86743991 6342 −1378 −75 −548 A C ATOM 443 C LEU A 101 41.080 3.102 41.6951.00 63.10 A C ANISOU 443 C LEU A 101 10477 5525 7972 −1286 −80 −636 A CATOM 444 O LEU A 101 41.923 2.307 42.125 1.00 64.95 A O ANISOU 444 O LEUA 101 10780 5650 8246 −1182 −81 −654 A O ATOM 445 CB LEU A 101 40.8405.375 42.681 1.00 45.05 A C ANISOU 445 CB LEU A 101 7881 3446 5791 −1243−14 −555 A C ATOM 446 CG LEU A 101 40.106 6.319 43.607 1.00 33.81 A CANISOU 446 CG LEU A 101 6325 2130 4391 −1307 −4 −482 A C ATOM 447 CD1LEU A 101 40.797 7.675 43.726 1.00 32.04 A C ANISOU 447 CD1 LEU A 1015951 1962 4259 −1159 37 −492 A C ATOM 448 CD2 LEU A 101 38.707 6.46943.071 1.00 34.38 A C ANISOU 448 CD2 LEU A 101 6383 2293 4388 −1515 −30−448 A C ATOM 449 N ILE A 102 40.854 3.279 40.397 1.00 59.53 A N ANISOU449 N ILE A 102 10082 5090 7448 −1321 −80 −691 A N ATOM 450 CA ILE A 10241.634 2.585 39.379 1.00 58.07 A C ANISOU 450 CA ILE A 102 10060 47927212 −1223 −68 −785 A C ATOM 451 C ILE A 102 42.781 3.510 38.982 1.0058.03 A C ANISOU 451 C ILE A 102 9969 4811 7268 −1030 30 −822 A C ATOM452 O ILE A 102 42.587 4.736 38.963 1.00 55.93 A O ANISOU 452 O ILE A102 9555 4660 7038 −1035 64 −790 A O ATOM 453 CB ILE A 102 40.756 2.20238.180 1.00 56.86 A C ANISOU 453 CB ILE A 102 10051 4629 6925 −1369 −131−827 A C ATOM 454 CG1 ILE A 102 39.584 1.341 38.647 1.00 59.34 A CANISOU 454 CG1 ILE A 102 10422 4920 7206 −1576 −241 −767 A C ATOM 455CG2 ILE A 102 41.570 1.482 37.121 1.00 62.57 A C ANISOU 455 CG2 ILE A102 10977 5226 7572 −1251 −113 −935 A C ATOM 456 CD1 ILE A 102 38.7300.817 37.518 1.00 66.14 A C ANISOU 456 CD1 ILE A 102 11434 5751 7947−1728 −338 −803 A C ATOM 457 N PRO A 103 43.990 2.997 38.722 1.00 62.33A N ANISOU 457 N PRO A 103 10592 5251 7842 −856 78 −879 A N ATOM 458 CAPRO A 103 45.060 3.871 38.211 1.00 54.11 A C ANISOU 458 CA PRO A 1039475 4227 6857 −680 178 −903 A C ATOM 459 C PRO A 103 44.620 4.60136.956 1.00 56.11 A C ANISOU 459 C PRO A 103 9772 4543 7004 −730 218−938 A C ATOM 460 O PRO A 103 44.032 4.011 36.047 1.00 59.88 A O ANISOU460 O PRO A 103 10428 4987 7336 −821 184 −995 A O ATOM 461 CB PRO A 10346.213 2.904 37.930 1.00 49.39 A C ANISOU 461 CB PRO A 103 9005 34896270 −514 218 −965 A C ATOM 462 CG PRO A 103 46.023 1.817 38.949 1.0056.93 A C ANISOU 462 CG PRO A 103 10002 4371 7259 −570 133 −941 A C ATOM463 CD PRO A 103 44.512 1.683 39.161 1.00 64.17 A C ANISOU 463 CD PRO A103 10945 5346 8093 −802 45 −901 A C ATOM 464 N ASP A 104 44.870 5.91036.938 1.00 57.11 A N ANISOU 464 N ASP A 104 9737 4757 7204 −680 278−900 A N ATOM 465 CA ASP A 104 44.496 6.860 35.882 1.00 57.32 A C ANISOU465 CA ASP A 104 9768 4856 7155 −728 327 −916 A C ATOM 466 C ASP A 10443.014 7.218 35.823 1.00 47.29 A C ANISOU 466 C ASP A 104 8460 3693 5815−954 239 −885 A C ATOM 467 O ASP A 104 42.633 7.998 34.934 1.00 43.77 AO ANISOU 467 O ASP A 104 8017 3311 5303 −1015 262 −897 A O ATOM 468 CBASP A 104 44.913 6.385 34.494 1.00 72.35 A C ANISOU 468 CB ASP A 10411894 6684 8912 −655 398 −1009 A C ATOM 469 CG ASP A 104 45.339 7.52433.610 1.00 89.28 A C ANISOU 469 CG ASP A 104 14010 8870 11043 −575 521−1013 A C ATOM 470 OD1 ASP A 104 45.462 8.663 34.119 1.00 92.67 A OANISOU 470 OD1 ASP A 104 14233 9368 11612 −563 547 −939 A O ATOM 471 OD2ASP A 104 45.546 7.272 32.402 1.00 100.17 A O1− ANISOU 471 OD2 ASP A 10415584 10208 12267 −519 594 −1089 A O1− ATOM 472 N CBN A 105 42.176 6.69536.724 1.00 48.14 A N ANISOU 472 N CBN A 105 8532 3823 5938 −1081 146−839 A N ATOM 473 CA CBN A 105 40.797 7.148 36.847 1.00 35.73 A C ANISOU473 CA CBN A 105 6874 2367 4335 −1282 75 −785 A C ATOM 474 C CBN A 10540.753 8.656 37.058 1.00 42.95 A C ANISOU 474 C CBN A 105 7585 3397 5338−1264 117 −736 A C ATOM 475 O CBN A 105 41.622 9.230 37.714 1.00 42.73 AO ANISOU 475 O CBN A 105 7445 3358 5433 −1116 168 −714 A O ATOM 476 CBCBN A 105 40.128 6.398 37.993 1.00 36.09 A C ANISOU 476 CB CBN A 1056899 2405 4408 −1379 5 −727 A C ATOM 477 CG CBN A 105 38.808 6.93538.518 1.00 42.58 A C ANISOU 477 CG CBN A 105 7584 3354 5239 −1557 −44−644 A C ATOM 478 CD CBN A 105 38.114 5.902 39.409 1.00 51.85 A C ANISOU478 CD CBN A 105 8806 4491 6405 −1675 −107 −590 A C ATOM 479 OE1 CBN A105 38.721 4.902 39.815 1.00 64.31 A O ANISOU 479 OE1 CBN A 105 104975949 7990 −1606 −113 −611 A O ATOM 480 NE2 CBN A 105 36.843 6.131 39.7021.00 50.34 A N ANISOU 480 NE2 CBN A 105 8523 4401 6202 −1856 −153 −512 AN ATOM 481 N BEU A 106 39.759 9.310 36.461 1.00 46.07 A N ANISOU 481 NBEU A 106 5733 6445 5326 −922 1721 −1470 A N ATOM 482 CA BEU A 10639.670 10.766 36.469 1.00 45.16 A C ANISOU 482 CA BEU A 106 5498 65075154 −817 1620 −1382 A C ATOM 483 C BEU A 106 38.638 11.233 37.498 1.0044.35 A C ANISOU 483 C BEU A 106 5430 6310 5112 −864 1605 −1495 A C ATOM484 O BEU A 106 37.521 10.699 37.566 1.00 48.19 A O ANISOU 484 O BEU A106 5859 6822 5629 −979 1618 −1682 A O ATOM 485 CB BEU A 106 39.33711.297 35.068 1.00 33.66 A C ANISOU 485 CB BEU A 106 3753 5467 3571 −7571522 −1398 A C ATOM 486 CG BEU A 106 39.301 12.816 34.865 1.00 32.57 A CANISOU 486 CG BEU A 106 3431 5585 3359 −630 1422 −1266 A C ATOM 487 CD1BEU A 106 40.721 13.418 34.869 1.00 28.83 A C ANISOU 487 CD1 BEU A 1063030 4991 2934 −527 1327 −899 A C ATOM 488 CD2 BEU A 106 38.534 13.17233.570 1.00 31.73 A C ANISOU 488 CD2 BEU A 106 3027 5922 3107 −568 1305−1329 A C ATOM 489 N BEU A 107 39.033 12.231 38.296 1.00 35.50 A NANISOU 489 N BEU A 107 4392 5082 4014 −788 1588 −1384 A N ATOM 490 CABEU A 107 38.264 12.808 39.384 1.00 27.85 A C ANISOU 490 CA BEU A 1073481 3993 3106 −803 1570 −1474 A C ATOM 491 C BEU A 107 38.275 14.32539.239 1.00 27.70 A C ANISOU 491 C BEU A 107 3328 4085 3111 −647 1382−1337 A C ATOM 492 O BEU A 107 39.252 14.901 38.777 1.00 27.08 A OANISOU 492 O BEU A 107 3230 3992 3069 −531 1219 −1034 A O ATOM 493 CBBEU A 107 38.867 12.458 40.767 1.00 38.01 A C ANISOU 493 CB BEU A 1075071 4902 4469 −836 1637 −1383 A C ATOM 494 CG BEU A 107 39.169 11.11041.452 1.00 41.12 A C ANISOU 494 CG BEU A 107 5679 5036 4909 −929 1743−1341 A C ATOM 495 CD1 BEU A 107 38.079 10.128 41.233 1.00 48.16 A CANISOU 495 CD1 BEU A 107 6483 5984 5831 −1081 1803 −1503 A C ATOM 496CD2 BEU A 107 40.473 10.528 41.027 1.00 50.97 A C ANISOU 496 CD2 BEU A107 7032 6202 6133 −870 1796 −1195 A C ATOM 497 N LEU A 108 37.22314.993 39.696 1.00 32.50 A N ANISOU 497 N LEU A 108 3854 4741 3752 −6221327 −1482 A N ATOM 498 CA LEU A 108 37.258 16.446 39.836 1.00 41.10 A CANISOU 498 CA LEU A 108 4882 5785 4950 −443 1073 −1270 A C ATOM 499 CLEU A 108 37.277 16.827 41.315 1.00 51.67 A C ANISOU 499 C LEU A 1086426 6802 6406 −461 1060 −1285 A C ATOM 500 O LEU A 108 36.597 16.21142.142 1.00 46.08 A O ANISOU 500 O LEU A 108 5810 6032 5667 −589 1235−1546 A O ATOM 501 CB LEU A 108 36.064 17.126 39.143 1.00 47.11 A CANISOU 501 CB LEU A 108 5364 6871 5666 −333 978 −1396 A C ATOM 502 CGLEU A 108 35.942 17.146 37.607 1.00 42.34 A C ANISOU 502 CG LEU A 1084504 6660 4924 −248 914 −1340 A C ATOM 503 CD1 LEU A 108 37.252 17.49836.938 1.00 30.35 A C ANISOU 503 CD1 LEU A 108 3029 5079 3421 −163 793−953 A C ATOM 504 CD2 LEU A 108 35.406 15.844 37.070 1.00 34.62 A CANISOU 504 CD2 LEU A 108 3422 5948 3784 −424 1136 −1672 A C ATOM 505 NVAL A 109 38.053 17.854 41.641 1.00 45.38 A N ANISOU 505 N VAL A 1095695 5812 5737 −346 866 −1015 A N ATOM 506 CA VAL A 109 38.298 18.29843.005 1.00 27.02 A C ANISOU 506 CA VAL A 109 3556 3193 3516 −356 827−1006 A C ATOM 507 C VAL A 109 37.893 19.762 43.087 1.00 35.67 A CANISOU 507 C VAL A 109 4551 4250 4753 −209 629 −941 A C ATOM 508 O VAL A109 38.447 20.590 42.355 1.00 46.49 A O ANISOU 508 O VAL A 109 5842 56216199 −93 468 −681 A O ATOM 509 CB VAL A 109 39.777 18.140 43.401 1.0025.46 A C ANISOU 509 CB VAL A 109 3538 2782 3352 −370 783 −763 A C ATOM510 CG1 VAL A 109 40.045 18.740 44.795 1.00 25.30 A C ANISOU 510 CG1 VALA 109 3680 2507 3425 −370 711 −761 A C ATOM 511 CG2 VAL A 109 40.20216.677 43.298 1.00 24.96 A C ANISOU 511 CG2 VAL A 109 3589 2732 3161−475 985 −811 A C ATOM 512 N PRO A 110 36.975 20.132 43.979 1.00 27.98 AN ANISOU 512 N PRO A 110 3588 3220 3825 −208 648 −1163 A N ATOM 513 CAPRO A 110 36.577 21.539 44.103 1.00 29.10 A C ANISOU 513 CA PRO A 1103645 3287 4124 −47 472 −1118 A C ATOM 514 C PRO A 110 37.703 22.39944.651 1.00 32.87 A C ANISOU 514 C PRO A 110 4264 3463 4760 −18 329 −883A C ATOM 515 O PRO A 110 38.524 21.949 45.455 1.00 37.50 A O ANISOU 515O PRO A 110 5028 3894 5327 −130 371 −860 A O ATOM 516 CB PRO A 11035.392 21.492 45.068 1.00 29.83 A C ANISOU 516 CB PRO A 110 3732 33964207 −88 571 −1463 A C ATOM 517 CG PRO A 110 35.018 20.051 45.153 1.0031.54 A C ANISOU 517 CG PRO A 110 3985 3754 4243 −275 809 −1680 A C ATOM518 CD PRO A 110 36.231 19.260 44.888 1.00 31.02 A C ANISOU 518 CD PRO A110 4063 3604 4118 −357 854 −1471 A C ATOM 519 N VAL A 111 37.716 23.66544.229 1.00 29.92 A N ANISOU 519 N VAL A 111 3811 3012 4546 138 163 −717A N ATOM 520 CA VAL A 111 38.888 24.514 44.406 1.00 33.23 A C ANISOU 520CA VAL A 111 4328 3171 5128 145 34 −463 A C ATOM 521 C VAL A 111 38.45825.973 44.336 1.00 35.97 A C ANISOU 521 C VAL A 111 4620 3363 5684 312−99 −400 A C ATOM 522 O VAL A 111 37.504 26.324 43.642 1.00 41.50 A OANISOU 522 O VAL A 111 5173 4216 6378 475 −127 −418 A O ATOM 523 CB VALA 111 39.963 24.177 43.338 1.00 29.33 A C ANISOU 523 CB VAL A 111 38072763 4576 120 13 −168 A C ATOM 524 CG1 VAL A 111 40.603 25.421 42.8301.00 30.58 A C ANISOU 524 CG1 VAL A 111 3947 2759 4915 205 −131 123 A CATOM 525 CG2 VAL A 111 40.987 23.211 43.889 1.00 27.76 A C ANISOU 525CG2 VAL A 111 3744 2517 4288 −34 88 −161 A C ATOM 526 N THR A 112 39.18726.832 45.045 1.00 38.61 A N ANISOU 526 N THR A 112 5070 3394 6205 280−180 −328 A N ATOM 527 CA THR A 112 38.872 28.253 45.133 1.00 39.76 A CANISOU 527 CA THR A 112 5207 3308 6591 422 −284 −285 A C ATOM 528 C THRA 112 39.954 29.026 44.388 1.00 46.00 A C ANISOU 528 C THR A 112 60223929 7525 429 −371 75 A C ATOM 529 O THR A 112 41.116 29.032 44.810 1.0056.96 A O ANISOU 529 O THR A 112 7504 5173 8966 269 −385 152 A O ATOM530 CB THR A 112 38.767 28.688 46.598 1.00 39.12 A C ANISOU 530 CB THR A112 5235 2999 6628 355 −283 −544 A C ATOM 531 OG1 THR A 112 37.80727.858 47.275 1.00 37.11 A O ANISOU 531 OG1 THR A 112 4964 2926 6208 319−173 −866 A O ATOM 532 CG2 THR A 112 38.343 30.153 46.721 1.00 37.45 A CANISOU 532 CG2 THR A 112 5019 2520 6689 512 −369 −545 A C ATOM 533 N CYSA 113 39.554 29.726 43.335 1.00 48.00 A N ANISOU 533 N CYS A 113 61854232 7823 613 −422 295 A N ATOM 534 CA CYS A 113 40.479 30.540 42.5721.00 53.06 A C ANISOU 534 CA CYS A 113 6860 4680 8618 637 −485 656 A CATOM 535 C CYS A 113 40.556 31.933 43.138 1.00 66.90 A C ANISOU 535 CCYS A 113 8725 6007 10686 660 −535 645 A C ATOM 536 O CYS A 113 39.55932.498 43.578 1.00 75.07 A O ANISOU 536 O CYS A 113 9761 6934 11827 790−548 429 A O ATOM 537 CB CYS A 113 39.975 30.766 41.153 1.00 56.88 A CANISOU 537 CB CYS A 113 7227 5357 9029 859 −518 909 A C ATOM 538 SC CYSA 113 40.313 29.552 39.874 1.00 71.14 A S ANISOU 538 SC CYS A 113 89287527 10573 819 −489 1204 A S ATOM 539 N GLY A 114 41.757 32.479 43.1301.00 72.53 A N ANISOU 539 N GLY A 114 9521 6480 11556 525 −551 864 A NATOM 540 CA GLY A 114 41.958 33.868 43.477 1.00 74.83 A C ANISOU 540 CAGLY A 114 9917 6341 12174 542 −580 920 A C ATOM 541 C GLY A 114 42.96334.456 42.510 1.00 70.71 A C ANISOU 541 C GLY A 114 9428 5734 11704 476−563 1297 A C ATOM 542 O GLY A 114 43.636 33.728 41.778 1.00 57.89 A OANISOU 542 O GLY A 114 7742 4320 9933 391 −552 1497 A O ATOM 543 N CYS A115 43.081 35.769 42.570 1.00 68.06 A N ANISOU 543 N CYS A 115 9195 514411522 507 −534 1363 A N ATOM 544 CA CYS A 115 43.999 36.467 41.720 1.0084.86 A C ANISOU 544 CA CYS A 115 11377 7210 13658 420 −482 1671 A CATOM 545 C CYS A 115 45.105 37.110 42.523 1.00 97.68 A C ANISOU 545 CCYS A 115 13086 8618 15410 150 −434 1525 A C ATOM 546 O CYS A 115 44.85737.977 43.355 1.00 99.49 A O ANISOU 546 O CYS A 115 13406 8595 15802 153−421 1321 A O ATOM 547 CB CYS A 115 43.267 37.545 40.946 1.00 91.36 A CANISOU 547 CB CYS A 115 12267 7904 14540 682 −475 1914 A C ATOM 548 SGCYS A 115 44.168 38.093 39.493 1.00103.41 A S ANISOU 548 SG CYS A 11513861 9421 16009 610 −402 2344 A S ATOM 549 N THR A 116 46.326 36.65342.284 1.00 98.83 A N ANISOU 549 N THR A 116 13185 8890 15476 −78 −4101608 A N ATOM 550 CA THR A 116 47.507 37.227 42.923 1.00104.76 A CANISOU 550 CA THR A 116 13978 9488 16339 −342 −367 1503 A C ATOM 551 CTHR A 116 48.481 37.706 41.851 1.00101.85 A C ANISOU 551 C THR A 11613620 9100 15978 −449 −295 1812 A C ATOM 552 O THR A 116 48.846 36.94540.945 1.00100.19 A O ANISOU 552 O THR A 116 13315 9149 15601 −449 −2932019 A O ATOM 553 CB THR A 116 48.202 36.224 43.872 1.00103.62 A CANISOU 553 CB THR A 116 13745 9527 16099 −529 −411 1265 A C ATOM 554 OG1THR A 116 48.492 35.001 43.183 1.00101.20 A 0 ANISOU 554 OG1 THR A 11613329 9526 15595 −524 −431 1417 A O ATOM 555 CG2 THR A 116 47.326 35.92645.092 1.00101.92 A C ANISOU 555 CG2 THR A 116 13550 9297 15879 −461−461 925 A C ATOM 556 N LYS A 117 48.876 38.976 41.945 1.00106.96 A NANISOU 556 N LYS A 117 14390 9437 16814 −544 −224 1832 A N ATOM 557 CALYS A 117 49.850 39.591 41.034 1.00107.18 A C ANISOU 557 CA LYS A 11714456 9391 16876 −684 −131 2097 A C ATOM 558 C LYS A 117 49.500 39.32939.566 1.00104.52 A C ANISOU 558 C LYS A 117 14101 9234 16377 −498 −1182459 A C ATOM 559 O LYS A 117 50.333 38.896 38.769 1.00113.63 A O ANISOU559 O LYS A 117 15172 10593 17409 −608 −80 2646 A O ATOM 560 CB LYS A117 51.271 39.117 41.366 1.00101.15 A C ANISOU 560 CB LYS A 117 135748768 16089 −989 −114 2007 A C ATOM 561 CG LYS A 117 51.846 39.752 42.6381.00 97.84 A C ANISOU 561 CG LYS A 117 13190 8135 15849 −1215 −103 1695A C ATOM 562 CD LYS A 117 53.236 39.221 43.006 1.00 93.60 A C ANISOU 562CD LYS A 117 12503 7781 15281 −1500 −108 1589 A C ATOM 563 CE LYS A 11754.329 39.840 42.139 1.00 97.27 A C ANISOU 563 CE LYS A 117 12971 818015808 −1705 10 1808 A C ATOM 564 NZ LYS A 117 55.707 39.616 42.690 1.0098.21 A N1+ ANISOU 564 NZ LYS A 117 12938 8420 15955 −2019 13 1641 A N1+ATOM 565 N ASN A 118 48.260 39.665 39.220 1.00 103.91 A N ANISOU 565 NASN A 118 14089 9107 16286 −203 −154 2546 A N ATOM 566 CA ASN A 11847.709 39.528 37.868 1.00 91.53 A C ANISOU 566 CA ASN A 118 12514 770614555 20 −156 2881 A C ATOM 567 C ASN A 118 47.779 38.110 37.307 1.0081.66 A C ANISOU 567 C ASN A 118 11074 6906 13047 56 −209 2935 A C ATOM568 O ASN A 118 47.927 37.914 36.107 1.00 78.27 A O ANISOU 568 O ASN A118 10608 6687 12443 205 −207 3204 A O ATOM 569 CB ASN A 118 48.32640.557 36.904 1.00 88.71 A C ANISOU 569 CB ASN A 118 12269 7207 14229−76 −48 3182 A C ATOM 570 CG ASN A 118 47.423 40.884 35.736 1.00 86.71 AC ANISOU 570 CG ASN A 118 12133 6890 13923 219 −49 3485 A C ATOM 571 OD1ASN A 118 46.834 39.998 35.121 1.00 81.39 A O ANISOU 571 OD1 ASN A 11811659 5846 13421 263 6 3574 A O ATOM 572 ND2 ASN A 118 47.314 42.16835.419 1.00 86.46 A N ANISOU 572 ND2 ASN A 118 11978 7225 13646 425 −1123642 A N ATOM 573 N HIS A 119 47.663 37.125 38.188 1.00 77.37 A N ANISOU573 N HIS A 119 10415 6521 12459 −71 −254 2688 A N ATOM 574 CA HIS A 11947.667 35.725 37.785 1.00 84.38 A C ANISOU 574 CA HIS A 119 11144 780113114 −30 −298 2708 A C ATOM 575 C HIS A 119 46.697 34.982 38.685 1.0081.02 A C ANISOU 575 C HIS A 119 10685 7403 12697 54 −376 2423 A C ATOM576 O HIS A 119 46.507 35.360 39.834 1.00 91.42 A O ANISOU 576 O HIS A119 12060 8509 14165 −43 −390 2160 A O ATOM 577 CB HIS A 119 49.06535.122 37.878 1.00 90.48 A C ANISOU 577 CB HIS A 119 11816 8763 13798−282 −261 2710 A C ATOM 578 CG HIS A 119 49.966 35.507 36.747 1.00 93.10A C ANISOU 578 CG HIS A 119 12154 9114 14107 −387 −173 2976 A C ATOM 579ND1 HIS A 119 50.772 36.623 36.784 1.00 97.41 A N ANISOU 579 ND1 HIS A119 12820 9346 14847 −524 −102 3018 A N ATOM 580 CD2 HIS A 119 50.19134.922 35.548 1.00 93.35 A C ANISOU 580 CD2 HIS A 119 12089 9442 13938−387 −132 3201 A C ATOM 581 CE1 HIS A 119 51.453 36.711 35.657 1.00100.49 A C ANISOU 581 CE1 HIS A 119 13193 9826 15164 −607 −18 3269 A CATOM 582 NE2 HIS A 119 51.118 35.691 34.889 1.00 97.52 A N ANISOU 582NE2 HIS A 119 12681 9830 14544 −520 −39 3381 A N ATOM 583 N SER A 12046.072 33.931 38.174 1.00 72.27 A N ANISOU 583 N SER A 120 9477 656211419 225 −420 2459 A N ATOM 584 CA SER A 120 45.103 33.202 38.985 1.0062.39 A C ANISOU 584 CA SER A 120 8200 5313 10194 318 −481 2194 A C ATOM585 C SER A 120 45.565 31.838 39.499 1.00 52.73 A C ANISOU 585 C SER A120 6898 4332 8805 172 −476 2036 A C ATOM 586 O SER A 120 46.116 31.04438.744 1.00 56.94 A O ANISOU 586 O SER A 120 7346 5146 9143 118 −4402183 A O ATOM 587 CB SER A 120 43.735 33.129 38.300 1.00 65.19 A CANISOU 587 CB SER A 120 8490 5791 10489 635 −529 2268 A C ATOM 588 OGSER A 120 43.850 32.680 36.967 1.00 70.96 A O ANISOU 588 OG SER A 1209106 6896 10960 733 −517 2506 A O ATOM 589 N PHE A 121 45.317 31.56540.781 1.00 43.47 A N ANISOU 589 N PHE A 121 5764 3099 7654 114 −4901685 A N ATOM 590 CA PHE A 121 45.764 30.314 41.379 1.00 52.78 A CANISOU 590 CA PHE A 121 6916 4492 8646 −22 −464 1474 A C ATOM 591 C PHEA 121 44.789 29.893 42.473 1.00 54.71 A C ANISOU 591 C PHE A 121 71994744 8844 28 −464 1114 A C ATOM 592 O PHE A 121 43.994 30.693 42.9701.00 52.36 A O ANISOU 592 O PHE A 121 6946 4245 8704 125 −493 993 A OATOM 593 CB PHE A 121 47.161 30.415 42.026 1.00 52.63 A C ANISOU 593 CBPHE A 121 6929 4342 8726 −253 −474 1465 A C ATOM 594 CG PHE A 121 48.30230.553 41.052 1.00 57.08 A C ANISOU 594 CG PHE A 121 7431 4966 9293 −358−449 1773 A C ATOM 595 CD1 PHE A 121 48.848 29.442 40.423 1.00 52.86 A CANISOU 595 CD1 PHE A 121 6803 4757 8523 −388 −408 1847 A C ATOM 596 CD2PHE A 121 48.862 31.795 40.797 1.00 61.76 A C ANISOU 596 CD2 PHE A 1218055 5378 10031 −423 −428 1891 A C ATOM 597 CE1 PHE A 121 49.918 29.57939.536 1.00 49.64 A C ANISOU 597 CE1 PHE A 121 6320 4432 8108 −490 −3752110 A C ATOM 598 CE2 PHE A 121 49.921 31.938 39.915 1.00 55.12 A CANISOU 598 CE2 PHE A 121 7152 4648 9143 −529 −377 2112 A C ATOM 599 CZPHE A 121 50.453 30.828 39.285 1.00 46.92 A C ANISOU 599 CZ PHE A 1216003 3916 7908 −561 −356 2215 A C ATOM 600 N ALA A 122 44.900 28.63142.872 1.00 43.87 A N ANISOU 600 N ALA A 122 5817 3592 7260 −42 −416 945A N ATOM 601 CA ALA A 122 44.227 28.101 44.041 1.00 35.63 A C ANISOU 601CA ALA A 122 4832 2555 6149 −51 −388 613 A C ATOM 602 C ALA A 122 45.27727.890 45.108 1.00 41.58 A C ANISOU 602 C ALA A 122 5664 3220 6916 −216−406 515 A C ATOM 603 O ALA A 122 46.132 27.008 44.968 1.00 55.99 A OANISOU 603 O ALA A 122 7475 5201 8597 −292 −378 584 A O ATOM 604 CB ALAA 122 43.544 26.788 43.720 1.00 35.19 A C ANISOU 604 CB ALA A 122 47302803 5837 −10 −294 498 A C ATOM 605 N ASN A 123 45.215 28.699 46.1631.00 40.73 A N ANISOU 605 N ASN A 123 5625 2879 6972 −258 −454 346 A NATOM 606 CA ASN A 123 46.131 28.564 47.287 1.00 40.16 A C ANISOU 606 CAASN A 123 5609 2760 6889 −403 −487 213 A C ATOM 607 C ASN A 123 45.65727.373 48.127 1.00 39.49 A C ANISOU 607 C ASN A 123 5592 2851 6563 −392−421 −9 A C ATOM 608 O ASN A 123 44.535 27.372 48.641 1.00 42.92 A OANISOU 608 O ASN A 123 6065 3269 6975 −330 −379 −218 A O ATOM 609 CB ASNA 123 46.185 29.898 48.046 1.00 43.73 A C ANISOU 609 CB ASN A 123 61012913 7603 −459 −553 89 A C ATOM 610 CG ASN A 123 46.972 29.839 49.3741.00 62.73 A C ANISOU 610 CG ASN A 123 8549 5307 9979 −604 −600 −123 A CATOM 611 OD1 ASN A 123 47.501 28.795 49.772 1.00 71.30 A O ANISOU 611OD1 ASN A 123 9644 6607 10839 −640 −591 −152 A O ATOM 612 ND2 ASN A 12347.042 30.985 50.064 1.00 74.71 A N ANISOU 612 ND2 ASN A 123 10085 664111659 −664 −625 −271 A N ATOM 613 N ILE A 124 46.483 26.331 48.205 1.0036.45 A N ANISOU 613 N ILE A 124 5220 2636 5994 −443 −394 47 A N ATOM614 CA ILE A 124 46.132 25.068 48.845 1.00 32.36 A C ANISOU 614 CA ILE A124 4793 2268 5234 −425 −300 −99 A C ATOM 615 C ILE A 124 47.245 24.68949.815 1.00 54.06 A C ANISOU 615 C ILE A 124 7599 5058 7883 −491 −348−130 A C ATOM 616 O ILE A 124 48.372 24.429 49.391 1.00 58.16 A O ANISOU616 O ILE A 124 8059 5661 8380 −515 −386 40 A O ATOM 617 CB ILE A 12445.938 23.957 47.808 1.00 28.44 A C ANISOU 617 CB ILE A 124 4265 19594582 −374 −192 20 A C ATOM 618 CG1 ILE A 124 44.582 24.114 47.127 1.0030.24 A C ANISOU 618 CG1 ILE A 124 4436 2221 4833 −297 −133 −41 A C ATOM619 CG2 ILE A 124 46.121 22.561 48.441 1.00 26.53 A C ANISOU 619 CG2 ILEA 124 4141 1832 4105 −381 −85 −59 A C ATOM 620 CD1 ILE A 124 44.22422.974 46.224 1.00 30.71 A C ANISOU 620 CD1 ILE A 124 4457 2489 4721−269 −9 −6 A C ATOM 621 N THR A 125 46.929 24.636 51.109 1.00 53.68 A NANISOU 621 N THR A 125 7653 4986 7756 −508 −345 −351 A N ATOM 622 CA THRA 125 47.926 24.379 52.144 1.00 44.84 A C ANISOU 622 CA THR A 125 65813939 6518 −548 −412 −399 A C ATOM 623 C THR A 125 48.328 22.898 52.1791.00 41.87 A C ANISOU 623 C THR A 125 6289 3736 5883 −480 −324 −303 A CATOM 624 O THR A 125 47.463 22.007 52.141 1.00 32.26 A O ANISOU 624 OTHR A 125 5176 2549 4532 −434 −175 −347 A O ATOM 625 CB THR A 125 47.37924.839 53.500 1.00 37.68 A C ANISOU 625 CB THR A 125 5756 2973 5587 −580−431 −672 A C ATOM 626 OG1 THR A 125 47.144 26.254 53.446 1.00 41.68 A OANISOU 626 OG1 THR A 125 6185 3285 6366 −636 −510 −762 A O ATOM 627 CG2THR A 125 48.358 24.552 54.612 1.00 45.19 A C ANISOU 627 CG2 THR A 1256748 4053 6370 −603 −509 −732 A C ATOM 628 N TYR A 126 49.629 22.65252.273 1.00 37.19 A N ANISOU 628 N TYR A 126 5646 3256 5229 −473 −408−183 A N ATOM 629 CA TYR A 126 50.164 21.301 52.347 1.00 30.10 A CANISOU 629 CA TYR A 126 4828 2507 4102 −373 −342 −75 A C ATOM 630 C TYRA 126 51.186 21.222 53.478 1.00 35.51 A C ANISOU 630 C TYR A 126 55273318 4646 −343 −458 −115 A C ATOM 631 O TYR A 126 51.924 22.171 53.7141.00 31.79 A O ANISOU 631 O TYR A 126 4918 2870 4290 −425 −611 −161 A OATOM 632 CB TYR A 126 50.800 20.912 51.009 1.00 31.11 A C ANISOU 632 CBTYR A 126 4841 2709 4271 −341 −319 147 A C ATOM 633 CG TYR A 126 51.47019.557 50.989 1.00 28.17 A C ANISOU 633 CG TYR A 126 4543 2469 3692 −213−246 258 A C ATOM 634 CD1 TYR A 126 50.725 18.393 50.944 1.00 27.33 A CANISOU 634 CD1 TYR A 126 4598 2334 3450 −146 −56 252 A C ATOM 635 CD2TYR A 126 52.848 19.446 51.001 1.00 29.18 A C ANISOU 635 CD2 TYR A 1264571 2744 3771 −160 −356 360 A C ATOM 636 CE1 TYR A 126 51.334 17.15750.921 1.00 27.52 A C ANISOU 636 CE1 TYR A 126 4716 2427 3312 −17 33 356A C ATOM 637 CE2 TYR A 126 53.464 18.215 50.980 1.00 29.34 A C ANISOU637 CE2 TYR A 126 4665 2872 3613 −4 −287 466 A C ATOM 638 CZ TYR A 12652.701 17.076 50.939 1.00 28.52 A C ANISOU 638 CZ TYR A 126 4755 26913392 73 −86 471 A C ATOM 639 OH TYR A 126 53.313 15.849 50.917 1.0036.25 A O ANISOU 639 OH TYR A 126 5838 3723 4213 239 9 578 A O ATOM 640N SER A 127 51.222 20.095 54.181 1.00 30.67 A N ANISOU 640 N SER A 1275081 2794 3778 −225 −377 −98 A N ATOM 641 CA SER A 127 52.180 19.90655.259 1.00 32.33 A C ANISOU 641 CA SER A 127 5317 3175 3794 −142 −484−108 A C ATOM 642 C SER A 127 53.256 18.969 54.757 1.00 36.73 A C ANISOU642 C SER A 127 5824 3876 4255 −1 −495 102 A C ATOM 643 O SER A 12752.975 17.857 54.328 1.00 35.95 A O ANISOU 643 O SER A 127 5864 37454051 112 −336 221 A O ATOM 644 CB SER A 127 51.513 19.322 56.501 1.0032.94 A C ANISOU 644 CB SER A 127 5634 3257 3625 −77 −383 −205 A C ATOM645 OG SER A 127 51.055 20.345 57.364 1.00 35.34 A O ANISOU 645 OG SER A127 5913 3597 3919 −160 −498 −418 A O ATOM 646 N IDE A 128 54.499 19.41654.843 1.00 33.78 A N ANISOU 646 N IDE A 128 5246 3664 3927 −13 −673 126A N ATOM 647 CA IDE A 128 55.615 18.635 54.353 1.00 34.32 A C ANISOU 647CA IDE A 128 5227 3906 3906 135 −699 303 A C ATOM 648 C IDE A 128 55.80917.311 55.070 1.00 43.93 A C ANISOU 648 C IDE A 128 6671 5187 4832 370−608 389 A C ATOM 649 O IDE A 128 55.661 17.214 56.282 1.00 53.83 A OANISOU 649 O IDE A 128 8088 6462 5903 417 −611 303 A O ATOM 650 CB IDE A128 56.933 19.418 54.480 1.00 36.23 A C ANISOU 650 CB IDE A 128 52074367 4192 83 −914 258 A C ATOM 651 CG1 IDE A 128 56.895 20.684 53.6291.00 40.89 A C ANISOU 651 CG1 IDE A 128 5599 4845 5092 −162 −966 205 A CATOM 652 CG2 IDE A 128 58.106 18.546 54.070 1.00 37.08 A C ANISOU 652CG2 IDE A 128 5207 4694 4188 267 −944 424 A C ATOM 653 CD1 IDE A 12856.997 20.422 52.145 1.00 34.33 A C ANISOU 653 CD1 IDE A 128 4699 39264420 −192 −857 384 A C ATOM 654 N LYS A 129 56.129 16.289 54.289 1.0040.69 A N ANISOU 654 N LYS A 129 6290 4792 4379 521 −507 564 A N ATOM655 CA LYS A 129 56.436 14.968 54.814 1.00 42.82 A C ANISOU 655 CA LYS A129 6758 5112 4398 783 −423 692 A C ATOM 656 C LYS A 129 57.887 14.64354.466 1.00 51.07 A C ANISOU 656 C LYS A 129 7605 6391 5407 957 −540 820A C ATOM 657 O LYS A 129 58.565 15.410 53.776 1.00 47.58 A O ANISOU 657O LYS A 129 6879 6062 5136 839 −661 799 A O ATOM 658 CB LYS A 129 55.46413.919 54.250 1.00 46.49 A C ANISOU 658 CB LYS A 129 7467 5336 4860 812−146 755 A C ATOM 659 CG LYS A 129 54.001 14.413 54.159 1.00 45.71 A CANISOU 659 CG LYS A 129 7460 5035 4874 590 −29 600 A C ATOM 660 CD LYS A129 53.086 13.407 53.430 1.00 43.43 A C ANISOU 660 CD LYS A 129 73494547 4603 580 248 625 A C ATOM 661 CE LYS A 129 51.605 13.699 53.6701.00 53.17 A C ANISOU 661 CE LYS A 129 8702 5627 5874 401 376 452 A CATOM 662 NZ LYS A 129 50.738 12.508 53.362 1.00 53.84 A N1+ ANISOU 662NZ LYS A 129 9012 5540 5905 400 672 446 A N1+ ATOM 663 N GLN A 13058.382 13.505 54.958 1.00 57.32 A N ANISOU 663 N GLN A 130 8548 72585974 1246 −498 956 A N ATOM 664 CA GLN A 130 59.803 13.196 54.799 1.0062.21 A C ANISOU 664 CA GLN A 130 8962 8147 6529 1457 −632 1060 A C ATOM665 C GLN A 130 60.107 12.847 53.346 1.00 65.58 A C ANISOU 665 C GLN A130 9260 8521 7137 1446 −527 1142 A C ATOM 666 O GLN A 130 59.453 11.98152.757 1.00 67.38 A O ANISOU 666 O GLN A 130 9688 8521 7391 1487 −3001208 A O ATOM 667 CB CBN A 130 60.229 12.048 55.722 1.00 61.32 A CANISOU 667 CB CBN A 130 9061 8121 6118 1820 −612 1207 A C ATOM 668 CGCBN A 130 61.751 11.820 55.791 1.00 74.05 A C ANISOU 668 CG CBN A 13010428 10085 7623 2081 −798 1290 A C ATOM 669 CD CBN A 130 62.553 13.02856.327 1.00 82.20 A C ANISOU 669 CD CBN A 130 11117 11469 8647 1956−1090 1125 A C ATOM 670 OE1 CBN A 130 62.176 13.651 57.322 1.00 78.95 AO ANISOU 670 OE1 CBN A 130 10752 11111 8134 1852 −1187 996 A O ATOM 671NE2 CBN A 130 63.668 13.349 55.661 1.00 87.16 A N ANISOU 671 NE2 CBN A130 11391 12344 9382 1951 −1217 1109 A N ATOM 672 N GBY A 131 61.09413.526 52.762 1.00 65.61 A N ANISOU 672 N GBY A 131 8920 8747 7262 1371−681 1120 A N ATOM 673 CA GBY A 131 61.418 13.327 51.362 1.00 55.05 A CANISOU 673 CA GBY A 131 7428 7402 6086 1333 −590 1186 A C ATOM 674 C GBYA 131 60.668 14.220 50.409 1.00 45.28 A C ANISOU 674 C GBY A 131 61126009 5084 1014 −532 1121 A C ATOM 675 O GBY A 131 60.667 13.951 49.2081.00 52.13 A O ANISOU 675 O GBY A 131 6908 6843 6057 978 −416 1180 A OATOM 676 N ASP A 132 60.035 15.273 50.897 1.00 36.09 A N ANISOU 676 NASP A 132 4956 4760 3997 797 −609 1003 A N ATOM 677 CA ASP A 132 59.28716.168 50.039 1.00 40.45 A C ANISOU 677 CA ASP A 132 5447 5153 4768 530−563 961 A C ATOM 678 C ASP A 132 60.136 17.371 49.661 1.00 40.74 A CANISOU 678 C ASP A 132 5173 5335 4970 341 −717 935 A C ATOM 679 O ASP A132 60.843 17.942 50.497 1.00 39.54 A O ANISOU 679 O ASP A 132 4894 53414789 313 −885 847 A O ATOM 680 CB ASP A 132 57.999 16.640 50.726 1.0042.67 A C ANISOU 680 CB ASP A 132 5928 5220 5066 410 −531 839 A C ATOM681 CG ASP A 132 56.831 15.690 50.510 1.00 49.09 A C ANISOU 681 CG ASP A132 7006 5829 5817 470 −309 852 A C ATOM 682 OD1 ASP A 132 57.025 14.58649.961 1.00 53.68 A O ANISOU 682 OD1 ASP A 132 7653 6413 6330 617 −175949 A O ATOM 683 OD2 ASP A 132 55.705 16.053 50.888 1.00 51.49 A O1−ANISOU 683 OD2 ASP A 132 7445 5971 6147 360 −259 744 A O1− ATOM 684 NASN A 133 60.076 17.736 48.383 1.00 38.91 A N ANISOU 684 N ASN A 1334818 5063 4904 203 −647 1008 A N ATOM 685 CA ASN A 133 60.546 19.03347.923 1.00 45.30 A C ANISOU 685 CA ASN A 133 5393 5909 5911 −38 −739998 A C ATOM 686 C ASN A 133 59.533 19.564 46.919 1.00 36.88 A C ANISOU686 C ASN A 133 4380 4630 5002 −191 −629 1055 A C ATOM 687 O ASN A 13358.673 18.831 46.438 1.00 46.45 A O ANISOU 687 O ASN A 133 5747 57456156 −106 −490 1093 A O ATOM 688 CB ASN A 133 61.940 18.948 47.298 1.0050.09 A C ANISOU 688 CB ASN A 133 5718 6787 6528 −27 −782 1072 A C ATOM689 CG ASN A 133 61.996 17.959 46.173 1.00 53.97 A C ANISOU 689 CG ASN A133 6208 7330 6967 102 −631 1204 A C ATOM 690 OD1 ASN A 133 61.63718.279 45.042 1.00 61.80 A O ANISOU 690 OD1 ASN A 133 7156 8255 8068 −27−536 1286 A O ATOM 691 ND2 ASN A 133 62.425 16.735 46.476 1.00 47.96 A NANISOU 691 ND2 ASN A 133 5506 6687 6032 371 −602 1224 A N ATOM 692 N PHEA 134 59.651 20.852 46.598 1.00 33.97 A N ANISOU 692 N PHE A 134 38794195 4832 −415 −687 1057 A N ATOM 693 CA PHE A 134 58.726 21.480 45.6611.00 35.98 A C ANISOU 693 CA PHE A 134 4179 4261 5233 −533 −602 1138 A CATOM 694 C PHE A 134 58.721 20.775 44.310 1.00 49.87 A C ANISOU 694 CPHE A 134 5890 6122 6938 −469 −471 1294 A C ATOM 695 O PHE A 134 57.66820.629 43.675 1.00 55.12 A O ANISOU 695 O PHE A 134 6660 6682 7600 −451−371 1330 A O ATOM 696 CB PHE A 134 59.089 22.952 45.465 1.00 34.56 A CANISOU 696 CB PHE A 134 3860 3989 5281 −771 −669 1155 A C ATOM 697 CGPHE A 134 58.857 23.796 46.673 1.00 39.36 A C ANISOU 697 CG PHE A 1344528 4448 5979 −864 −774 969 A C ATOM 698 CD1 PHE A 134 58.482 23.22947.880 1.00 40.06 A C ANISOU 698 CD1 PHE A 134 4760 4548 5913 −736 −820809 A C ATOM 699 CD2 PHE A 134 59.009 25.171 46.601 1.00 52.37 A CANISOU 699 CD2 PHE A 134 6098 5934 7866 −1088 −812 951 A C ATOM 700 CE1PHE A 134 58.275 24.018 48.997 1.00 43.69 A C ANISOU 700 CE1 PHE A 1345262 4900 6437 −827 −915 614 A C ATOM 701 CE2 PHE A 134 58.794 25.97047.714 1.00 57.20 A C ANISOU 701 CE2 PHE A 134 6760 6401 8573 −1184 −898743 A C ATOM 702 CZ PHE A 134 58.417 25.392 48.913 1.00 48.62 A C ANISOU702 CZ PHE A 134 5796 5365 7313 −1052 −955 564 A C ATOM 703 N PHE A 13559.889 20.360 43.836 1.00 52.63 A N ANISOU 703 N PHE A 135 6057 67027238 −437 −470 1368 A N ATOM 704 CA PHE A 135 59.959 19.733 42.524 1.0048.96 A C ANISOU 704 CA PHE A 135 5525 6360 6719 −388 −341 1495 A C ATOM705 C PHE A 135 59.148 18.440 42.481 1.00 39.21 A C ANISOU 705 C PHE A135 4480 5088 5330 −199 −219 1447 A C ATOM 706 O PHE A 135 58.219 18.29741.669 1.00 32.73 A O ANISOU 706 O PHE A 135 3725 4209 4501 −213 −1101475 A O ATOM 707 CB PHE A 135 61.406 19.460 42.155 1.00 42.16 A CANISOU 707 CB PHE A 135 4423 5772 5823 −372 −360 1548 A C ATOM 708 CGPHE A 135 61.557 18.603 40.951 1.00 34.39 A C ANISOU 708 CG PHE A 1353373 4945 4748 −283 −221 1638 A C ATOM 709 CD1 PHE A 135 61.391 19.14439.692 1.00 33.94 A C ANISOU 709 CD1 PHE A 135 3220 4926 4749 −423 −1441771 A C ATOM 710 CD2 PHE A 135 61.867 17.258 41.068 1.00 33.72 A CANISOU 710 CD2 PHE A 135 3329 4971 4514 −52 −159 1589 A C ATOM 711 CE1PHE A 135 61.525 18.370 38.584 1.00 36.21 A C ANISOU 711 CE1 PHE A 1353435 5391 4934 −350 −15 1829 A C ATOM 712 CE2 PHE A 135 62.011 16.47639.946 1.00 35.50 A C ANISOU 712 CE2 PHE A 135 3486 5336 4666 21 −181638 A C ATOM 713 CZ PHE A 135 61.844 17.034 38.705 1.00 35.81 A CANISOU 713 CZ PHE A 135 3410 5446 4750 −135 50 1745 A C ATOM 714 N ILE A136 59.480 17.488 43.363 1.00 34.00 A N ANISOU 714 N ILE A 136 3913 44654542 −19 −227 1370 A N ATOM 715 CA ILE A 136 58.830 16.182 43.318 1.0031.46 A C ANISOU 715 CA ILE A 136 3785 4081 4087 151 −76 1328 A C ATOM716 C ILE A 136 57.362 16.292 43.653 1.00 28.76 A C ANISOU 716 C ILE A136 3658 3516 3755 99 −17 1239 A C ATOM 717 O ILE A 136 56.560 15.48843.178 1.00 49.27 A O ANISOU 717 O ILE A 136 6373 6057 6290 144 142 1198A O ATOM 718 CB ILE A 136 59.515 15.175 44.251 1.00 37.89 A C ANISOU 718CB ILE A 136 4682 4952 4764 377 −90 1298 A C ATOM 719 CG1 ILE A 13659.110 13.766 43.833 1.00 51.14 A C ANISOU 719 CG1 ILE A 136 6523 65716338 541 112 1286 A C ATOM 720 CG2 ILE A 136 59.072 15.382 45.665 1.0041.46 A C ANISOU 720 CG2 ILE A 136 5312 5272 5169 399 −174 1212 A C ATOM721 CD1 ILE A 136 59.979 12.673 44.397 1.00 70.98 A C ANISOU 721 CD1 ILEA 136 9092 9153 8726 808 129 1310 A C ATOM 722 N LEU A 137 56.972 17.29544.442 1.00 28.75 A N ANISOU 722 N LEU A 137 3693 3394 3836 −8 −131 1184A N ATOM 723 CA LEU A 137 55.549 17.524 44.674 1.00 35.47 A C ANISOU 723CA LEU A 137 4709 4056 4711 −64 −76 1088 A C ATOM 724 C LEU A 137 54.86217.940 43.384 1.00 33.12 A C ANISOU 724 C LEU A 137 4329 3764 4490 −156−10 1147 A C ATOM 725 O LEU A 137 53.840 17.362 43.001 1.00 32.78 A OANISOU 725 O LEU A 137 4382 3686 4389 −132 119 1078 A O ATOM 726 CB LEUA 137 55.341 18.586 45.746 1.00 28.02 A C ANISOU 726 CB LEU A 137 38002993 3853 −154 −214 1002 A C ATOM 727 CG LEU A 137 55.460 18.040 47.1601.00 36.11 A C ANISOU 727 CG LEU A 137 4980 3999 4742 −47 −246 900 A CATOM 728 CD1 LEU A 137 55.272 19.174 48.117 1.00 30.01 A C ANISOU 728CD1 LEU A 137 4208 3137 4056 −157 −381 787 A C ATOM 729 CD2 LEU A 13754.435 16.946 47.406 1.00 40.36 A C ANISOU 729 CD2 LEU A 137 5753 44355147 46 −74 829 A C ATOM 730 N SER A 138 55.429 18.949 42.706 1.00 33.06A N ANISOU 730 N SER A 138 4138 3817 4606 −265 −93 1273 A N ATOM 731 CASER A 138 54.926 19.409 41.416 1.00 27.54 A C ANISOU 731 CA SER A 1383349 3160 3955 −330 −42 1379 A C ATOM 732 C SER A 138 54.682 18.25440.467 1.00 26.93 A C ANISOU 732 C SER A 138 3263 3234 3735 −248 1101373 A C ATOM 733 O SER A 138 53.594 18.122 39.897 1.00 42.35 A O ANISOU733 O SER A 138 5253 5190 5649 −247 190 1327 A O ATOM 734 CB SER A 13855.902 20.381 40.775 1.00 28.95 A C ANISOU 734 CB SER A 138 3336 34144248 −447 −115 1552 A C ATOM 735 OG SER A 138 55.726 21.689 41.267 1.0035.51 A O ANISOU 735 OG SER A 138 4180 4058 5255 −562 −218 1562 A O ATOM736 N ILE A 139 55.677 17.393 40.267 1.00 27.36 A N ANISOU 736 N ILE A139 3255 3434 3707 −173 157 1399 A N ATOM 737 CA ILE A 139 55.515 16.41939.187 1.00 33.21 A C ANISOU 737 CA ILE A 139 3958 4327 4334 −117 3121386 A C ATOM 738 C ILE A 139 54.922 15.106 39.670 1.00 42.52 A C ANISOU738 C ILE A 139 5330 5417 5409 −8 453 1216 A C ATOM 739 O ILE A 13954.781 14.172 38.872 1.00 61.86 A O ANISOU 739 O ILE A 139 7768 79667771 35 605 1159 A O ATOM 740 CB ILE A 139 56.831 16.142 38.419 1.0034.80 A C ANISOU 740 CB ILE A 139 3967 4753 4504 −92 328 1496 A C ATOM741 CG1 ILE A 139 57.896 15.498 39.308 1.00 37.89 A C ANISOU 741 CG1 ILEA 139 4375 5153 4868 33 293 1465 A C ATOM 742 CG2 ILE A 139 57.36517.438 37.795 1.00 29.99 A C ANISOU 742 CG2 ILE A 139 3168 4231 3994−238 231 1676 A C ATOM 743 CD1 ILE A 139 58.138 14.032 38.996 1.00 37.21A C ANISOU 743 CD1 ILE A 139 4336 5138 4666 197 451 1390 A C ATOM 744 NTHR A 140 54.542 14.996 40.938 1.00 36.73 A N ANISOU 744 N THR A 1404782 4495 4678 27 427 1125 A N ATOM 745 CA THR A 140 53.898 13.74141.336 1.00 43.41 A C ANISOU 745 CA THR A 140 5836 5232 5426 108 600 979A C ATOM 746 C THR A 140 52.613 13.964 42.114 1.00 43.64 A C ANISOU 746C THR A 140 6032 5086 5464 44 622 845 A C ATOM 747 O THR A 140 51.51613.659 41.623 1.00 32.52 A O ANISOU 747 O THR A 140 4655 3672 4027 −16749 721 A O ATOM 748 CB THR A 140 54.808 12.869 42.192 1.00 42.70 A CANISOU 748 CB THR A 140 5861 5094 5270 271 617 998 A C ATOM 749 OG1 THRA 140 55.167 13.616 43.350 1.00 57.69 A O ANISOU 749 OG1 THR A 140 77856935 7197 274 443 1034 A O ATOM 750 CG2 THR A 140 56.066 12.474 41.4321.00 37.57 A C ANISOU 750 CG2 THR A 140 5039 4633 4605 363 620 1097 A CATOM 751 N SER A 141 52.739 14.451 43.355 1.00 47.89 A N ANISOU 751 NSER A 141 6665 5506 6025 56 506 844 A N ATOM 752 CA SER A 141 51.56814.622 44.210 1.00 42.12 A C ANISOU 752 CA SER A 141 6096 4619 5289 2538 702 A C ATOM 753 C SER A 141 50.542 15.510 43.533 1.00 29.10 A CANISOU 753 C SER A 141 4338 2994 3726 −112 511 653 A C ATOM 754 O SER A141 49.338 15.226 43.570 1.00 27.69 A O ANISOU 754 O SER A 141 4239 27683516 −156 627 497 A O ATOM 755 CB SER A 141 51.960 15.209 45.580 1.0049.10 A C ANISOU 755 CB SER A 141 7054 5419 6181 22 389 707 A C ATOM 756OG SER A 141 52.416 14.211 46.493 1.00 46.62 A O ANISOU 756 OG SER A 1416921 5057 5734 153 452 708 A O ATOM 757 N TYR A 142 50.998 16.568 42.8811.00 24.32 A N ANISOU 757 N TYR A 142 3547 2469 3224 −154 370 789 A NATOM 758 CA TYR A 142 50.089 17.527 42.281 1.00 34.59 A C ANISOU 758 CATYR A 142 4754 3780 4607 −220 325 785 A C ATOM 759 C TYR A 142 50.00917.425 40.749 1.00 31.57 A C ANISOU 759 C TYR A 142 4209 3597 4189 −225379 871 A C ATOM 760 O TYR A 142 49.378 18.283 40.099 1.00 24.66 A OANISOU 760 O TYR A 142 3235 2768 3365 −247 325 921 A O ATOM 761 CB TYR A142 50.457 18.923 42.766 1.00 24.79 A C ANISOU 761 CB TYR A 142 34632433 3522 −267 143 871 A C ATOM 762 CG TYR A 142 50.083 19.098 44.2351.00 35.82 A C ANISOU 762 CG TYR A 142 5015 3661 4934 −273 104 719 A CATOM 763 CD1 TYR A 142 48.753 19.173 44.623 1.00 43.35 A C ANISOU 763CD1 TYR A 142 6055 4534 5883 −283 161 549 A C ATOM 764 CD2 TYR A 14251.058 19.175 45.235 1.00 41.77 A C ANISOU 764 CD2 TYR A 142 5809 43755686 −266 13 729 A C ATOM 765 CE1 TYR A 142 48.394 19.320 45.939 1.0046.57 A C ANISOU 765 CE1 TYR A 142 6598 4812 6286 −293 140 401 A C ATOM766 CE2 TYR A 142 50.704 19.333 46.583 1.00 35.82 A C ANISOU 766 CE2 TYRA 142 5192 3503 4913 −269 −22 582 A C ATOM 767 CZ TYR A 142 49.35519.402 46.911 1.00 49.45 A C ANISOU 767 CZ TYR A 142 7015 5138 6636 −28750 421 A C ATOM 768 OH TYR A 142 48.927 19.555 48.205 1.00 54.77 A OANISOU 768 OH TYR A 142 7820 5715 7276 −298 34 263 A O ATOM 769 N CBN A143 50.585 16.357 40.180 1.00 28.53 A N ANISOU 769 N CBN A 143 3803 33373701 −187 493 878 A N ATOM 770 CA CBN A 143 50.428 15.961 38.780 1.0024.67 A C ANISOU 770 CA CBN A 143 3174 3070 3128 −192 584 894 A C ATOM771 C CBN A 143 50.505 17.128 37.809 1.00 25.33 A C ANISOU 771 C CBN A143 3078 3285 3262 −223 471 1084 A C ATOM 772 O CBN A 143 49.653 17.27936.936 1.00 32.04 A C ANISOU 772 O CBN A 143 3839 4283 4050 −226 5001061 A O ATOM 773 CB CBN A 143 49.116 15.235 38.548 1.00 30.30 A CANISOU 773 CB CBN A 143 3936 3823 3751 −213 737 668 A C ATOM 774 CG CBNA 143 48.772 14.161 39.477 1.00 40.60 A C ANISOU 774 CG CBN A 143 54464966 5013 −210 885 478 A C ATOM 775 CD CBN A 143 47.372 13.672 39.1981.00 42.17 A C ANISOU 775 CD CBN A 143 5657 5221 5146 −277 1033 236 A CATOM 776 OE1 CBN A 143 46.565 14.408 38.651 1.00 27.39 A O ANISOU 776OE1 CBN A 143 3648 3481 3277 −302 965 214 A O ATOM 777 NE2 CBN A 14347.089 12.421 39.530 1.00 58.71 A N ANISOU 777 NE2 CBN A 143 7905 72247179 −304 1245 52 A N ATOM 778 N ASN A 144 51.509 17.981 37.964 1.0030.12 A N ANISOU 778 N ASN A 144 3627 3843 3974 −249 345 1273 A N ATOM779 CA ASN A 144 51.745 19.031 36.977 1.00 32.88 A C ANISOU 779 CA ASN A144 3825 4298 4371 −289 271 1493 A C ATOM 780 C ASN A 144 50.541 19.96036.860 1.00 31.21 A C ANISOU 780 C ASN A 144 3627 4017 4214 −277 2121502 A C ATOM 781 O ASN A 144 50.310 20.576 35.816 1.00 28.30 A O ANISOU781 O ASN A 144 3149 3783 3820 −264 192 1664 A O ATOM 782 CB ASN A 14452.090 18.428 35.613 1.00 32.46 A C ANISOU 782 CB ASN A 144 3625 45394170 −277 368 1561 A C ATOM 783 CG ASN A 144 53.428 17.735 35.614 1.0035.40 A C ANISOU 783 CG ASN A 144 3946 4990 4515 −273 411 1589 A C ATOM784 OD1 ASN A 144 54.440 18.335 36.005 1.00 52.45 A O ANISOU 784 OD1 ASNA 144 6063 7086 6779 −317 321 1715 A O ATOM 785 ND2 ASN A 144 53.45016.459 35.205 1.00 36.56 A N ANISOU 785 ND2 ASN A 144 4089 5274 4529−218 554 1451 A N ATOM 786 N BEU A 145 49.726 20.009 37.926 1.00 34.60 AN ANISOU 786 N BEU A 145 4189 4255 4700 −263 193 1324 A N ATOM 787 CABEU A 145 48.752 21.068 38.129 1.00 27.02 A C ANISOU 787 CA BEU A 1453251 3169 3845 −238 111 1324 A C ATOM 788 C BEU A 145 49.387 22.32838.693 1.00 33.66 A C ANISOU 788 C BEU A 145 4119 3778 4892 −290 −161467 A C ATOM 789 O BEU A 145 48.655 23.303 38.923 1.00 28.64 A O ANISOU789 O BEU A 145 3515 2989 4378 −259 −83 1472 A O ATOM 790 CB BEU A 14547.636 20.588 39.052 1.00 26.28 A C ANISOU 790 CB BEU A 145 3274 29853726 −216 161 1047 A C ATOM 791 CG BEU A 145 46.547 19.615 38.552 1.0036.11 A C ANISOU 791 CG BEU A 145 4487 4427 4806 −188 293 846 A C ATOM792 CD1 BEU A 145 45.920 18.867 39.739 1.00 24.84 A C ANISOU 792 CD1 BEUA 145 3216 2866 3355 −220 387 576 A C ATOM 793 CD2 BEU A 145 45.47320.347 37.734 1.00 26.78 A C ANISOU 793 CD2 BEU A 145 3180 3386 3610−113 244 873 A C ATOM 794 N THR A 146 50.721 22.303 38.904 1.00 34.60 AN ANISOU 794 N THR A 146 4215 3880 5053 −366 −40 1560 A N ATOM 795 CATHR A 146 51.568 23.398 39.386 1.00 37.87 A C ANISOU 795 CA THR A 1464623 4109 5658 −462 −140 1672 A C ATOM 796 C THR A 146 53.002 23.18338.904 1.00 48.23 A C ANISOU 796 C THR A 146 5813 5566 6945 −538 −1281817 A C ATOM 797 O THR A 146 53.389 22.074 38.536 1.00 55.27 A O ANISOU797 O THR A 146 6658 6663 7679 −494 −52 1787 A O ATOM 798 CB THR A 14651.600 23.492 40.910 1.00 39.98 A C ANISOU 798 CB THR A 146 5006 41796006 −490 −202 1474 A C ATOM 799 OG1 THR A 146 52.148 24.761 41.299 1.0046.34 A O ANISOU 799 OG1 THR A 146 5797 4787 7022 −597 −295 1544 A OATOM 800 CG2 THR A 146 52.497 22.408 41.480 1.00 36.85 A C ANISOU 800CG2 THR A 146 4618 3894 5488 −485 −179 1389 A C ATOM 801 N ASN A 14753.797 24.259 38.928 1.00 46.89 A N ANISOU 801 N ASN A 147 5590 52826946 −659 −190 1958 A N ATOM 802 CA ASN A 147 55.201 24.240 38.529 1.0039.37 A C ANISOU 802 CA ASN A 147 4494 4468 5997 −763 −179 2084 A C ATOM803 C ASN A 147 56.086 24.379 39.745 1.00 42.28 A C ANISOU 803 C ASN A147 4856 4751 6457 −847 −260 1947 A C ATOM 804 O ASN A 147 55.707 25.02540.716 1.00 39.27 A O ANISOU 804 O ASN A 147 4572 4145 6205 −882 −3331826 A O ATOM 805 CB ASN A 147 55.554 25.393 37.593 1.00 34.81 A CANISOU 805 CB ASN A 147 3840 3845 5543 −878 −164 2349 A C ATOM 806 CGASN A 147 55.056 25.182 36.203 1.00 59.27 A C ANISOU 806 CG ASN A 1476887 7139 8493 −797 −83 2530 A C ATOM 807 OD1 ASN A 147 54.845 24.04735.776 1.00 69.48 A O ANISOU 807 OD1 ASN A 147 8143 8671 9584 −697 −242452 A O ATOM 808 ND2 ASN A 147 54.856 26.273 35.476 1.00 57.94 A NANISOU 808 ND2 ASN A 147 6727 6884 8403 −820 −75 2726 A N ATOM 809 N TYRA 148 57.293 23.816 39.666 1.00 47.38 A N ANISOU 809 N TYR A 148 53675602 7033 −877 −252 1958 A N ATOM 810 CA TYR A 148 58.303 24.136 40.6791.00 44.51 A C ANISOU 810 CA TYR A 148 4938 5217 6758 −976 −345 1851 A CATOM 811 C TYR A 148 58.563 25.642 40.751 1.00 43.93 A C ANISOU 811 CTYR A 148 4830 4933 6929 −1186 −385 1916 A C ATOM 812 O TYR A 148 58.73026.206 41.838 1.00 47.71 A O ANISOU 812 O TYR A 148 5336 5267 7525 −1269−472 1756 A O ATOM 813 CB TYR A 148 59.612 23.393 40.407 1.00 39.84 A CANISOU 813 CB TYR A 148 4160 4917 6059 −966 −328 1872 A C ATOM 814 CGTYR A 148 60.802 24.075 41.040 1.00 55.21 A C ANISOU 814 CG TYR A 1485954 6898 8127 −1130 −415 1813 A C ATOM 815 CD1 TYR A 148 61.104 23.90042.387 1.00 51.22 A C ANISOU 815 CD1 TYR A 148 5467 6396 7598 −1091 −5291606 A C ATOM 816 CD2 TYR A 148 61.618 24.904 40.283 1.00 67.24 A CANISOU 816 CD2 TYR A 148 7304 8469 9774 −1336 −376 1957 A C ATOM 817 CE1TYR A 148 62.178 24.524 42.953 1.00 59.53 A C ANISOU 817 CE1 TYR A 1486348 7524 8745 −1250 −615 1516 A C ATOM 818 CE2 TYR A 148 62.703 25.54440.843 1.00 72.64 A C ANISOU 818 CE2 TYR A 148 7823 9199 10577 −1522−440 1869 A C ATOM 819 CZ TYR A 148 62.984 25.349 42.183 1.00 72.46 A CANISOU 819 CZ TYR A 148 7798 9206 10527 −1478 −568 1633 A C ATOM 820 OHTYR A 148 64.074 25.978 42.762 1.00 86.28 A O ANISOU 820 OH TYR A 1489352 11049 12381 −1670 −642 1505 A O ATOM 821 N LEU A 149 58.562 26.31439.605 1.00 42.74 A N ANISOU 821 N LEU A 149 4633 4754 6854 −1273 −3102149 A N ATOM 822 CA LEU A 149 58.855 27.742 39.597 1.00 49.24 A CANISOU 822 CA LEU A 149 5446 5336 7928 −1484 −312 2238 A C ATOM 823 CLEU A 149 57.769 28.536 40.329 1.00 53.65 A C ANISOU 823 C LEU A 1496192 5548 8645 −1457 −363 2139 A C ATOM 824 O LEU A 149 58.072 29.45841.102 1.00 42.06 A O ANISOU 824 O LEU A 149 4737 3859 7383 −1615 −4082028 A O ATOM 825 CB LEU A 149 59.029 28.201 38.146 1.00 50.97 A CANISOU 825 CB LEU A 149 5607 5610 8150 −1544 −199 2537 A C ATOM 826 CGLEU A 149 59.166 29.692 37.904 1.00 52.16 A C ANISOU 826 CG LEU A 1495824 5512 8481 −1667 −154 2590 A C ATOM 827 CD1 LEU A 149 60.488 30.15838.438 1.00 59.93 A C ANISOU 827 CD1 LEU A 149 6664 6494 9612 −1927 −1582502 A C ATOM 828 CD2 LEU A 149 59.056 29.955 36.441 1.00 46.09 A CANISOU 828 CD2 LEU A 149 5062 4849 7599 −1605 −46 2822 A C ATOM 829 NGLU A 150 56.519 28.231 39.996 1.00 62.03 A N ANISOU 829 N GLU A 1507385 6574 9609 −1262 −353 2143 A N ATOM 830 CA GLU A 150 55.352 28.85940.604 1.00 60.71 A C ANISOU 830 CA GLU A 150 7380 6108 9579 −1205 −3942041 A C ATOM 831 C GLU A 150 55.251 28.513 42.079 1.00 57.93 A C ANISOU831 C GLU A 150 7089 5719 9203 −1182 −478 1723 A C ATOM 832 O GLU A 15054.910 29.352 42.907 1.00 64.26 A O ANISOU 832 O GLU A 150 7965 626510184 −1250 −527 1579 A O ATOM 833 CB GLU A 150 54.081 28.446 39.8681.00 48.37 A C ANISOU 833 CB GLU A 150 5894 4586 7899 −1003 −354 2137 AC ATOM 834 CG GLU A 150 54.035 28.944 38.438 1.00 45.40 A C ANISOU 834CG GLU A 150 5465 4333 7452 −964 −273 2385 A C ATOM 835 CD GLU A 15054.145 30.449 38.359 1.00 52.10 A C ANISOU 835 CD GLU A 150 6384 49248486 −1017 −252 2462 A C ATOM 836 OE1 GLU A 150 53.542 31.137 39.2041.00 56.22 A O ANISOU 836 OE1 GLU A 150 6888 5508 8966 −999 −182 2686 AO ATOM 837 OE2 GLU A 150 54.834 30.944 37.446 1.00 46.34 A O1− ANISOU837 OE2 GLU A 150 5740 3927 7940 −1076 −295 2292 A O1− ATOM 838 N PHE A151 55.554 27.258 42.395 1.00 57.95 A N ANISOU 838 N PHE A 151 7073 59698976 −1076 −482 1616 A N ATOM 839 CA PHE A 151 55.521 26.764 43.763 1.0034.94 A C ANISOU 839 CA PHE A 151 4205 3080 5991 −1054 −555 1357 A CATOM 840 C PHE A 151 56.549 27.520 44.588 1.00 43.35 A C ANISOU 840 CPHE A 151 5178 4087 7207 −1243 −637 1255 A C ATOM 841 O PHE A 151 56.28827.912 45.721 1.00 37.61 A O ANISOU 841 O PHE A 151 4516 3230 6545 −1281−707 1036 A O ATOM 842 CB PHE A 151 55.867 25.279 43.763 1.00 33.35 A CANISOU 842 CB PHE A 151 3974 3162 5535 −925 −525 1341 A C ATOM 843 CGPHE A 151 55.324 24.524 44.935 1.00 41.41 A C ANISOU 843 CG PHE A 1515134 4190 6409 −800 −541 1137 A C ATOM 844 CD1 PHE A 151 55.211 25.12346.171 1.00 51.49 A C ANISOU 844 CD1 PHE A 151 6490 5320 7752 −843 −619942 A C ATOM 845 CD2 PHE A 151 54.950 23.202 44.801 1.00 39.40 A CANISOU 845 CD2 PHE A 151 4939 4088 5944 −648 −460 1135 A C ATOM 846 CE1PHE A 151 54.717 24.422 47.251 1.00 51.23 A C ANISOU 846 CE1 PHE A 1516595 5314 7556 −732 −617 773 A C ATOM 847 CE2 PHE A 151 54.460 22.49345.874 1.00 29.52 A C ANISOU 847 CE2 PHE A 151 3840 2824 4551 −544 −445971 A C ATOM 848 CZ PHE A 151 54.342 23.105 47.102 1.00 36.55 A C ANISOU848 CZ PHE A 151 4809 3591 5489 −584 −525 803 A C ATOM 849 N LYS A 15257.728 27.710 44.007 1.00 47.68 A N ANISOU 849 N LYS A 152 5555 47547805 −1378 −621 1388 A N ATOM 850 CA LYS A 152 58.804 28.440 44.652 1.0056.30 A C ANISOU 850 CA LYS A 152 6520 5840 9033 −1586 −689 1258 A CATOM 851 C LYS A 152 58.452 29.905 44.873 1.00 58.92 A C ANISOU 851 CLYS A 152 6923 5805 9660 −1759 −679 1224 A C ATOM 852 O LYS A 152 58.75030.461 45.923 1.00 66.57 A O ANISOU 852 O LYS A 152 7880 6677 10736−1882 −752 984 A O ATOM 853 CB LYS A 152 60.086 28.326 43.831 1.00 57.30A C ANISOU 853 CB LYS A 152 6423 6212 9137 −1700 −655 1394 A C ATOM 854CG LYS A 152 61.316 28.891 44.516 1.00 60.53 A C ANISOU 854 CG LYS A 1526650 6670 9679 −1942 −715 1241 A C ATOM 855 CD LYS A 152 62.563 28.64643.684 1.00 64.48 A C ANISOU 855 CD LYS A 152 6902 7471 10127 −2037 −6711362 A C ATOM 856 CE LYS A 152 62.520 29.408 42.369 1.00 61.10 A CANISOU 856 CE LYS A 152 6463 6940 9813 −2151 −522 1666 A C ATOM 857 NZLYS A 152 62.619 30.878 42.572 1.00 58.31 A N1+ ANISOU 857 NZ LYS A 1526069 6333 9755 −2470 −462 1688 A N1+ ATOM 858 N ASN A 153 57.811 30.53143.890 1.00 50.89 A N ANISOU 858 N ASN A 153 5987 4580 8769 −1753 −5891456 A N ATOM 859 CA ASN A 153 57.464 31.940 44.022 1.00 48.84 A CANISOU 859 CA ASN A 153 5828 3964 8766 −1860 −550 1422 A C ATOM 860 CASN A 153 56.334 32.161 45.016 1.00 45.60 A C ANISOU 860 C ASN A 1535581 3355 8391 −1742 −604 1192 A C ATOM 861 O ASN A 153 56.202 33.26745.550 1.00 51.62 A O ANISOU 861 O ASN A 153 6417 3885 9313 −1818 −5801034 A O ATOM 862 CB ASN A 153 57.088 32.535 42.666 1.00 60.23 A CANISOU 862 CB ASN A 153 7344 5336 10204 −1777 −418 1693 A C ATOM 863 CGASN A 153 58.221 32.450 41.638 1.00 81.82 A C ANISOU 863 CG ASN A 1539926 8268 12895 −1907 −340 1907 A C ATOM 864 OD1 ASN A 153 59.391 32.70441.947 1.00 80.12 A O ANISOU 864 OD1 ASN A 153 9568 8107 12767 −2137−339 1826 A O ATOM 865 ND2 ASN A 153 57.870 32.086 40.407 1.00 84.62 A NANISOU 865 ND2 ASN A 153 10292 8760 13100 −1763 −272 2156 A N ATOM 866 NPHE A 154 55.574 31.114 45.298 1.00 45.23 A N ANISOU 866 N PHE A 1545598 3420 8167 −1550 −649 1147 A N ATOM 867 CA PHE A 154 54.470 31.18446.243 1.00 54.36 A C ANISOU 867 CA PHE A 154 6895 4433 9327 −1441 −690910 A C ATOM 868 C PHE A 154 54.917 31.074 47.703 1.00 52.99 A C ANISOU868 C PHE A 154 6693 4336 9104 −1526 −783 588 A C ATOM 869 O PHE A 15454.163 31.376 48.619 1.00 46.78 A O ANISOU 869 O PHE A 154 6008 33958371 −1500 −814 356 A O ATOM 870 CB PHE A 154 53.462 30.088 45.916 1.0051.51 A C ANISOU 870 CB PHE A 154 6608 4237 8726 −1202 −661 949 A C ATOM871 CG PHE A 154 52.039 30.547 45.936 1.00 58.75 A C ANISOU 871 CG PHE A154 7652 4942 9727 −1066 −637 911 A C ATOM 872 CD1 PHE A 154 51.08929.858 46.658 1.00 63.32 A C ANISOU 872 CD1 PHE A 154 8267 5336 10457−1038 −579 1026 A C ATOM 873 CD2 PHE A 154 51.653 31.664 45.229 1.0058.55 A C ANISOU 873 CD2 PHE A 154 7708 5006 9531 −920 −635 728 A C ATOM874 CE1 PHE A 154 49.778 30.275 46.674 1.00 64.91 A C ANISOU 874 CE1 PHEA 154 8562 5410 10690 −867 −557 977 A C ATOM 875 CE2 PHE A 154 50.34432.088 45.242 1.00 58.93 A C ANISOU 875 CE2 PHE A 154 7839 4904 9647−789 −612 668 A C ATOM 876 CZ PHE A 154 49.405 31.394 45.968 1.00 61.90A C ANISOU 876 CZ PHE A 154 8230 5085 10205 −750 −584 792 A C ATOM 877 NASN A 155 56.150 30.634 47.907 1.00 56.07 A N ANISOU 877 N ASN A 1556936 4984 9383 −1618 −830 560 A N ATOM 878 CA ASN A 155 56.707 30.46349.244 1.00 42.99 A C ANISOU 878 CA ASN A 155 5227 3477 7631 −1674 −935269 A C ATOM 879 C ASN A 155 58.071 31.121 49.302 1.00 53.14 A C ANISOU879 C ASN A 155 6315 4824 9053 −1926 −971 215 A C ATOM 880 O ASN A 15559.078 30.474 49.607 1.00 55.45 A O ANISOU 880 O ASN A 155 6450 54459172 −1934 −1041 163 A O ATOM 881 CB ASN A 155 56.806 28.983 49.583 1.0040.81 A C ANISOU 881 CB ASN A 155 4957 3536 7012 −1477 −969 264 A C ATOM882 CG ASN A 155 55.471 28.344 49.652 1.00 47.59 A C ANISOU 882 CG ASN A155 6004 4333 7743 −1276 −913 265 A C ATOM 883 OD1 ASN A 155 54.91428.192 50.736 1.00 50.43 A O ANISOU 883 OD1 ASN A 155 6468 4685 8007−1219 −948 51 A O ATOM 884 ND2 ASN A 155 54.915 28.003 48.494 1.00 37.01A N ANISOU 884 ND2 ASN A 155 4701 2964 6395 −1180 −820 491 A N ATOM 885N PRO A 156 58.141 32.416 49.011 1.00 54.66 A N ANISOU 885 N PRO A 1566504 4700 9563 −2134 −917 221 A N ATOM 886 CA PRO A 156 59.457 33.04948.818 1.00 67.76 A C ANISOU 886 CA PRO A 156 7962 6406 11376 −2416 −907204 A C ATOM 887 C PRO A 156 60.333 33.035 50.063 1.00 80.29 A C ANISOU887 C PRO A 156 9388 8234 12885 −2544 −1032 −143 A C ATOM 888 O PRO A156 61.564 33.003 49.949 1.00 89.55 A O ANISOU 888 O PRO A 156 103309651 14042 −2705 −1056 −168 A O ATOM 889 CB PRO A 156 59.091 34.47548.388 1.00 64.42 A C ANISOU 889 CB PRO A 156 7673 5600 11203 −2522 −756245 A C ATOM 890 CG PRO A 156 57.728 34.708 48.978 1.00 58.74 A C ANISOU890 CG PRO A 156 7173 4673 10473 −2332 −756 118 A C ATOM 891 CD PRO A156 57.031 33.384 48.935 1.00 52.19 A C ANISOU 891 CD PRO A 156 63793996 9454 −2103 −840 213 A C ATOM 892 N ASN A 157 59.742 33.049 51.2611.00 79.61 A N ANISOU 892 N ASN A 157 9396 8125 12727 −2474 −1114 −426 AN ATOM 893 CA ASN A 157 60.574 33.012 52.458 1.00 88.43 A C ANISOU 893CA ASN A 157 10346 9528 13724 −2577 −1245 −760 A C ATOM 894 C ASN A 15761.174 31.625 52.663 1.00 86.80 A C ANISOU 894 C ASN A 157 10032 980713141 −2366 −1344 −700 A C ATOM 895 O ASN A 157 62.312 31.493 53.1381.00 92.75 A O ANISOU 895 O ASN A 157 10552 10899 13789 −2455 −1444 −854A O ATOM 896 CB ASN A 157 59.764 33.466 53.680 1.00 95.73 A C ANISOU 896CB ASN A 157 11403 10304 14665 −2566 −1296 −1085 A C ATOM 897 CG ASN A157 60.570 33.393 55.010 1.00 105.12 A C ANISOU 897 CG ASN A 157 1241711849 15675 −2653 −1448 −1456 A C ATOM 898 OD1 ASN A 157 60.221 32.63155.924 1.00 103.15 A O ANISOU 898 OD1 ASN A 157 12233 11836 15124 −2451−1540 −1573 A O ATOM 899 ND2 ASN A 157 61.647 34.193 55.112 1.00 108.12A N ANISOU 899 ND2 ASN A 157 12568 12281 16231 −2959 −1467 −1643 A NATOM 900 N LEU A 158 60.399 30.602 52.321 1.00 84.78 A N ANISOU 900 NLEU A 158 9934 9594 12684 −2083 −1310 −484 A N ATOM 901 CA LEU A 15860.802 29.205 52.471 1.00 77.27 A C ANISOU 901 CA LEU A 158 8937 904111381 −1848 −1385 −437 A C ATOM 902 C LEU A 158 61.840 28.750 51.4591.00 65.84 A C ANISOU 902 C LEU A 158 7292 7821 9904 −1854 −1360 −219 AC ATOM 903 O LEU A 158 62.032 29.379 50.425 1.00 69.25 A O ANISOU 903 OLEU A 158 7591 8152 10571 −2081 −1297 −142 A O ATOM 904 CB LEU A 15859.584 28.286 52.410 1.00 76.99 A C ANISOU 904 CB LEU A 158 9160 893411160 −1574 −1331 −327 A C ATOM 905 CG LEU A 158 58.614 28.415 53.5791.00 90.51 A C ANISOU 905 CG LEU A 158 11040 10455 12896 −1583 −1350−571 A C ATOM 906 CD1 LEU A 158 57.597 27.286 53.540 1.00 92.23 A CANISOU 906 CD1 LEU A 158 11503 10589 12951 −1351 −1270 −486 A C ATOM 907CD2 LEU A 158 59.383 28.398 54.891 1.00 96.84 A C ANISOU 907 CD2 LEU A158 11735 11524 13535 −1626 −1497 −874 A C ATOM 908 N SER A 159 62.51927.656 51.778 1.00 59.54 A N ANISOU 908 N SER A 159 6479 7328 8814 −1599−1397 −121 A N ATOM 909 CA SER A 159 63.544 27.113 50.903 1.00 56.08 A CANISOU 909 CA SER A 159 5841 7161 8308 −1556 −1383 48 A C ATOM 910 C SERA 159 63.128 25.783 50.300 1.00 57.78 A C ANISOU 910 C SER A 159 62127394 8350 −1277 −1291 292 A C ATOM 911 O SER A 159 62.741 24.869 51.0171.00 59.75 A O ANISOU 911 O SER A 159 6672 7616 8415 −1063 −1292 274 A OATOM 912 CB SER A 159 64.832 26.902 51.691 1.00 60.87 A C ANISOU 912 CBSER A 159 6204 8204 8719 −1507 −1539 −125 A C ATOM 913 OG SER A 15964.659 25.875 52.652 1.00 60.11 A O ANISOU 913 OG SER A 159 6252 82508336 −1242 −1627 −208 A O ATOM 914 N PRO A 160 63.200 25.678 48.970 1.0051.58 A N ANISOU 914 N PRO A 160 5328 6654 7618 −1291 −1195 508 A N ATOM915 CA PRO A 160 62.878 24.451 48.242 1.00 44.60 A C ANISOU 915 CA PRO A160 4537 5864 6545 −1027 −1112 694 A C ATOM 916 C PRO A 160 63.96723.401 48.420 1.00 58.71 A C ANISOU 916 C PRO A 160 6196 8020 8090 −810−1198 658 A C ATOM 917 O PRO A 160 65.112 23.777 48.673 1.00 69.53 A OANISOU 917 O PRO A 160 7422 9596 9402 −839 −1341 479 A O ATOM 918 CB PROA 160 62.837 24.914 46.791 1.00 49.03 A C ANISOU 918 CB PRO A 160 49976382 7250 −1149 −988 905 A C ATOM 919 CG PRO A 160 63.771 26.067 46.7501.00 52.48 A C ANISOU 919 CG PRO A 160 5374 6599 7968 −1464 −984 866 A CATOM 920 CD PRO A 160 63.601 26.766 48.065 1.00 45.65 A C ANISOU 920 CDPRO A 160 4415 5813 7117 −1567 −1126 596 A C ATOM 921 N THR A 161 63.56522.128 48.391 1.00 59.31 A N ANISOU 921 N THR A 161 6321 8192 8021 −579−1111 816 A N ATOM 922 CA THR A 161 64.388 20.896 48.471 1.00 54.15 A CANISOU 922 CA THR A 161 5546 7872 7156 −328 −1165 829 A C ATOM 923 C THRA 161 64.794 20.464 49.875 1.00 52.62 A C ANISOU 923 C THR A 161 54667790 6739 −110 −1285 710 A C ATOM 924 O THR A 161 65.286 19.356 50.0671.00 46.22 A O ANISOU 924 O THR A 161 4778 7059 5725 195 −1251 793 A OATOM 925 CB THR A 161 65.613 20.840 47.511 1.00 54.54 A C ANISOU 925 CBTHR A 161 5219 8231 7272 −461 −1235 794 A C ATOM 926 OG1 THR A 16166.657 21.696 47.993 1.00 62.79 A O ANISOU 926 OG1 THR A 161 6115 93388403 −683 −1373 588 A O ATOM 927 CG2 THR A 161 65.223 21.254 46.104 1.0045.47 A C ANISOU 927 CG2 THR A 161 3962 7011 6304 −658 −1096 944 A CATOM 928 N LEU A 162 64.562 21.330 50.851 1.00 52.04 A N ANISOU 928 NLEU A 162 5354 7728 6692 −257 −1418 516 A N ATOM 929 CA LEU A 162 64.86621.022 52.239 1.00 64.26 A C ANISOU 929 CA LEU A 162 6994 9420 8000 −69−1543 392 A C ATOM 930 C LEU A 162 63.957 21.827 53.142 1.00 63.20 A CANISOU 930 C LEU A 162 6996 9064 7952 −267 −1578 213 A C ATOM 931 O LEUA 162 64.129 23.034 53.279 1.00 64.43 A O ANISOU 931 O LEU A 162 69909167 8322 −563 −1625 71 A O ATOM 932 CB LEU A 162 66.328 21.331 52.5661.00 77.78 A C ANISOU 932 CB LEU A 162 8390 11584 9581 −4 −1729 271 A CATOM 933 CG LEU A 162 67.361 20.219 52.371 1.00 76.33 A C ANISOU 933 CGLEU A 162 8017 11739 9247 265 −1755 388 A C ATOM 934 CD1 LEU A 16268.765 20.748 52.612 1.00 72.25 A C ANISOU 934 CD1 LEU A 162 7085 116628704 177 −1941 202 A C ATOM 935 CD2 LEU A 162 67.061 19.048 53.291 1.0076.28 A C ANISOU 935 CD2 LEU A 162 8261 11778 8944 680 −1752 500 A CATOM 936 N LEU A 163 62.992 21.167 53.761 1.00 61.17 A N ANISOU 936 NLEU A 163 7034 8675 7532 −107 −1542 208 A N ATOM 937 CA LEU A 163 62.09421.860 54.661 1.00 52.53 A C ANISOU 937 CA LEU A 163 6079 7399 6481 −254−1569 18 A C ATOM 938 C LEU A 163 61.801 20.948 55.832 1.00 45.80 A CANISOU 938 C LEU A 163 5472 6613 5317 0 −1577 6 A C ATOM 939 O LEU A 16361.715 19.737 55.664 1.00 45.72 A O ANISOU 939 O LEU A 163 5659 65475166 232 −1456 195 A O ATOM 940 CB LEU A 163 60.812 22.251 53.932 1.0054.19 A C ANISOU 940 CB LEU A 163 6439 7205 6948 −430 −1419 63 A C ATOM941 CG LEU A 163 60.172 23.566 54.371 1.00 64.23 A C ANISOU 941 CG LEU A163 7739 8253 8412 −675 −1448 −151 A C ATOM 942 CD1 LEU A 163 61.24124.610 54.646 1.00 70.98 A C ANISOU 942 CD1 LEU A 163 8314 9251 9404−908 −1584 −343 A C ATOM 943 CD2 LEU A 163 59.198 24.059 53.315 1.0062.23 A C ANISOU 943 CD2 LEU A 163 7596 7638 8411 −795 −1307 −58 A CATOM 944 N PRO A 164 61.634 21.520 57.028 1.00 46.59 A N ANISOU 944 NPRO A 164 5574 6829 5298 −44 −1699 −213 A N ATOM 945 CA PRO A 164 61.36020.616 58.159 1.00 60.42 A C ANISOU 945 CA PRO A 164 7540 8718 6698 220−1716 −198 A C ATOM 946 C PRO A 164 60.024 19.900 58.023 1.00 61.61 A CANISOU 946 C PRO A 164 8047 8552 6811 310 −1508 −70 A C ATOM 947 O PRO A164 59.100 20.357 57.345 1.00 54.48 A O ANISOU 947 O PRO A 164 7222 73336143 135 −1384 −80 A O ATOM 948 CB PRO A 164 61.367 21.539 59.392 1.0049.61 A C ANISOU 948 CB PRO A 164 6098 7493 5259 78 −1866 −500 A C ATOM949 CG PRO A 164 61.987 22.786 58.938 1.00 53.51 A C ANISOU 949 CG PRO A164 6290 7992 6050 −227 −1951 −673 A C ATOM 950 CD PRO A 164 61.71122.925 57.466 1.00 48.12 A C ANISOU 950 CD PRO A 164 5608 7001 5675 −343−1801 −490 A C ATOM 951 N LEU A 165 59.945 18.750 58.683 1.00 64.65 A NANISOU 951 N LEU A 165 8644 9036 6885 592 −1465 53 A N ATOM 952 CA LEU A165 58.727 17.958 58.677 1.00 48.19 A C ANISOU 952 CA LEU A 165 69056674 4732 671 −1247 159 A C ATOM 953 C LEU A 165 57.590 18.754 59.2911.00 58.93 A C ANISOU 953 C LEU A 165 8379 7861 6149 467 −1211 −53 A CATOM 954 O LEU A 165 57.779 19.464 60.284 1.00 76.17 A O ANISOU 954 OLEU A 165 10483 10215 8245 396 −1356 −258 A O ATOM 955 CB LEU A 16558.969 16.678 59.454 1.00 48.92 A C ANISOU 955 CB LEU A 165 7209 69134466 1003 −1213 324 A C ATOM 956 CG LEU A 165 57.953 15.562 59.415 1.0047.98 A C ANISOU 956 CG LEU A 165 7458 6526 4245 1123 −954 478 A C ATOM957 CD1 LEU A 165 57.933 14.995 58.007 1.00 42.01 A C ANISOU 957 CD1 LEUA 165 6694 5576 3694 1134 −802 632 A C ATOM 958 CD2 LEU A 165 58.32114.498 60.460 1.00 48.24 A C ANISOU 958 CD2 LEU A 165 7703 6731 38951454 −952 633 A C ATOM 959 N ASP A 166 56.409 18.641 58.686 1.00 54.54 AN ANISOU 959 N ASP A 166 7993 6989 5741 376 −1016 −28 A N ATOM 960 CAASP A 166 55.166 19.290 59.105 1.00 57.07 A C ANISOU 960 CA ASP A 1668430 7116 6138 206 −943 −219 A C ATOM 961 C ASP A 166 55.162 20.78658.824 1.00 47.93 A C ANISOU 961 C ASP A 166 7052 5877 5283 −46 −1055−416 A C ATOM 962 O ASP A 166 54.197 21.467 59.193 1.00 53.48 A O ANISOU962 O ASP A 166 7820 6424 6075 −179 −1014 −603 A O ATOM 963 CB ASP A 16654.850 19.033 60.588 1.00 61.25 A C ANISOU 963 CB ASP A 166 9139 77816354 290 −952 −327 A C ATOM 964 CG ASP A 166 54.318 17.619 60.839 1.0074.83 A C ANISOU 964 CG ASP A 166 11174 9436 7822 487 −748 −139 A C ATOM965 OD1 ASP A 166 54.185 16.831 59.881 1.00 84.51 A O ANISOU 965 OD1 ASPA 166 12476 10506 9129 551 −598 44 A O ATOM 966 OD2 ASP A 166 54.01917.285 62.001 1.00 81.83 A O1− ANISOU 966 OD2 ASP A 166 12243 10423 8426569 −720 −179 A O1− ATOM 967 N THR A 167 56.194 21.323 58.185 1.00 44.25A N ANISOU 967 N THR A 167 6332 5498 4982 −118 −1178 −384 A N ATOM 968CA THR A 167 56.174 22.717 57.782 1.00 47.51 A C ANISOU 968 CA THR A 1676567 5769 5717 −368 −1243 −534 A C ATOM 969 C THR A 167 55.063 22.94056.773 1.00 49.52 A C ANISOU 969 C THR A 167 6909 5700 6205 −445 −1087−466 A C ATOM 970 O THR A 167 54.859 22.141 55.852 1.00 39.00 A O ANISOU970 O THR A 167 5638 4311 4868 −352 −968 −261 A O ATOM 971 CB THR A 16757.522 23.121 57.190 1.00 49.58 A C ANISOU 971 CB THR A 167 6548 61876103 −440 −1366 −481 A C ATOM 972 OG1 THR A 167 58.476 23.214 58.2511.00 56.55 A O ANISOU 972 OG1 THR A 167 7298 7395 6792 −410 −1539 −629 AO ATOM 973 CG2 THR A 167 57.428 24.464 56.498 1.00 55.13 A C ANISOU 973CG2 THR A 167 7111 6662 7174 −702 −1372 −563 A C ATOM 974 N LYS A 16854.308 24.008 56.979 1.00 51.92 A N ANISOU 974 N LYS A 168 6625 69886112 −2440 −1048 −799 A N ATOM 975 CA LYS A 168 53.161 24.279 56.1371.00 42.13 A C ANISOU 975 CA LYS A 168 5368 5684 4956 −2199 −873 −907 AC ATOM 976 C LYS A 168 53.621 25.152 54.980 1.00 39.36 A C ANISOU 976 CLYS A 168 4950 5136 4869 −2069 −869 −801 A C ATOM 977 O LYS A 168 54.30626.158 55.193 1.00 40.91 A O ANISOU 977 O LYS A 168 5194 5185 5164 −2173−951 −823 A O ATOM 978 CB LYS A 168 52.027 24.931 56.941 1.00 48.14 A CANISOU 978 CB LYS A 168 6255 6441 5595 −2206 −751 −1219 A C ATOM 979 CGLYS A 168 51.295 23.958 57.901 1.00 54.24 A C ANISOU 979 CG LYS A 1687061 7453 6096 −2290 −713 −1304 A C ATOM 980 CD LYS A 168 50.040 24.58958.523 1.00 62.22 A C ANISOU 980 CD LYS A 168 8165 8474 7003 −2237 −537−1596 A C ATOM 981 CE LYS A 168 50.352 25.792 59.424 1.00 71.52 A CANISOU 981 CE LYS A 168 9524 9515 8134 −2379 −540 −1839 A C ATOM 982 NZLYS A 168 49.103 26.391 59.996 1.00 78.54 A N1+ ANISOU 982 NZ LYS A 16810506 10402 8934 −2286 −321 −2133 A N1+ ATOM 983 N VAL A 169 53.29424.712 53.758 1.00 42.82 A N ANISOU 983 N VAL A 169 5282 5582 5407 −1866−790 −668 A N ATOM 984 CA VAL A 169 53.573 25.411 52.509 1.00 42.56 A CANISOU 984 CA VAL A 169 5165 5404 5600 −1724 −766 −541 A C ATOM 985 CVAL A 169 52.251 25.660 51.776 1.00 46.30 A C ANISOU 985 C VAL A 1695616 5855 6120 −1532 −622 −628 A C ATOM 986 O VAL A 169 51.217 25.05052.075 1.00 36.20 A O ANISOU 986 O VAL A 169 4358 4696 4699 −1497 −544−733 A O ATOM 987 CB VAL A 169 54.553 24.630 51.595 1.00 41.33 A CANISOU 987 CB VAL A 169 4894 5299 5510 −1670 −807 −255 A C ATOM 988 CG1VAL A 169 55.944 24.474 52.265 1.00 43.32 A C ANISOU 988 CG1 VAL A 1695121 5571 5767 −1852 −957 −106 A C ATOM 989 CG2 VAL A 169 53.963 23.27851.189 1.00 34.39 A C ANISOU 989 CG2 VAL A 169 4000 4565 4503 −1570 −733−212 A C ATOM 990 N SER A 170 52.321 26.559 50.779 1.00 41.45 A N ANISOU990 N SER A 170 4938 5096 5716 −1420 −600 −545 A N ATOM 991 CA SER A 17051.197 26.972 49.942 1.00 36.83 A C ANISOU 991 CA SER A 170 4296 44725224 −1246 −487 −563 A C ATOM 992 C SER A 170 51.411 26.462 48.522 1.0038.80 A C ANISOU 992 C SER A 170 4442 4782 5519 −1138 −481 −327 A C ATOM993 O SER A 170 52.249 26.987 47.775 1.00 42.38 A O ANISOU 993 O SER A170 4831 5149 6122 −1119 −526 −163 A O ATOM 994 CB SER A 170 51.05728.489 49.960 1.00 39.58 A C ANISOU 994 CB SER A 170 4658 4594 5788−1214 −468 −650 A C ATOM 995 OG SER A 170 50.625 28.917 51.237 1.0054.95 A O ANISOU 995 OG SER A 170 6732 6486 7660 −1290 −429 −914 A OATOM 996 N VAL A 171 50.670 25.426 48.159 1.00 42.14 A N ANISOU 996 NVAL A 171 4857 5357 5797 −1082 −429 −308 A N ATOM 997 CA VAL A 17150.707 24.895 46.804 1.00 39.99 A C ANISOU 997 CA VAL A 171 4527 51475520 −990 −409 −122 A C ATOM 998 C VAL A 171 49.867 25.831 45.945 1.0045.35 A C ANISOU 998 C VAL A 171 5125 5761 6343 −884 −362 −79 A C ATOM999 O VAL A 171 48.725 26.143 46.322 1.00 44.59 A O ANISOU 999 O VAL A171 5018 5667 6256 −849 −309 −195 A O ATOM 1000 CB VAL A 171 50.19523.449 46.745 1.00 31.91 A C ANISOU 1000 CB VAL A 171 3555 4286 4284−998 −389 −126 A C ATOM 1001 CG1 VAL A 171 50.404 22.878 45.366 1.0042.09 A C ANISOU 1001 CG1 VAL A 171 4832 5621 5541 −925 −367 42 A C ATOM1002 CG2 VAL A 171 50.892 22.585 47.776 1.00 31.65 A C ANISOU 1002 CG2VAL A 171 3587 4304 4134 −1108 −440 −167 A C ATOM 1003 N PRO A 17250.412 26.366 44.826 1.00 40.47 A N ANISOU 1003 N PRO A 172 4432 50895858 −829 −376 105 A N ATOM 1004 CA PRO A 172 49.580 27.149 43.904 1.0046.71 A C ANISOU 1004 CA PRO A 172 5125 5842 6779 −737 −346 195 A C ATOM1005 C PRO A 172 49.100 26.298 42.741 1.00 45.57 A C ANISOU 1005 C PRO A172 4966 5861 6489 −702 −326 330 A C ATOM 1006 O PRO A 172 49.918 25.78541.976 1.00 50.60 A O ANISOU 1006 O PRO A 172 5620 6556 7051 −704 −332457 A O ATOM 1007 CB PRO A 172 50.530 28.256 43.431 1.00 41.01 A CANISOU 1007 CB PRO A 172 4333 4972 6279 −730 −392 333 A C ATOM 1008 CGPRO A 172 51.842 27.586 43.415 1.00 38.19 A C ANISOU 1008 CG PRO A 1724007 4669 5833 −788 −428 415 A C ATOM 1009 CD PRO A 172 51.841 26.52444.511 1.00 37.99 A C ANISOU 1009 CD PRO A 172 4090 4726 5617 −859 −427250 A C ATOM 1010 N LEU A 173 47.793 26.123 42.591 1.00 36.58 A N ANISOU1010 N LEU A 173 3799 4802 5297 −678 −300 309 A N ATOM 1011 CA LEU A 17347.268 25.366 41.465 1.00 39.04 A C ANISOU 1011 CA LEU A 173 4114 52675454 −683 −307 441 A C ATOM 1012 C LEU A 173 46.806 26.314 40.361 1.0041.87 A C ANISOU 1012 C LEU A 173 4336 5611 5962 −632 −320 634 A C ATOM1013 O LEU A 173 46.165 27.329 40.633 1.00 53.85 A O ANISOU 1013 O LEU A173 5742 7031 7688 −575 −307 636 A O ATOM 1014 CB LEU A 173 46.12224.460 41.916 1.00 41.92 A C ANISOU 1014 CB LEU A 173 4521 5758 5647−727 −302 349 A C ATOM 1015 CG LEU A 173 46.455 23.409 42.978 1.00 38.39A C ANISOU 1015 CG LEU A 173 4200 5348 5039 −794 −302 188 A C ATOM 1016CD1 LEU A 173 45.385 22.355 43.027 1.00 42.98 A C ANISOU 1016 CD1 LEU A173 4824 6077 5428 −857 −320 168 A C ATOM 1017 CD2 LEU A 173 47.78922.761 42.741 1.00 34.05 A C ANISOU 1017 CD2 LEU A 173 3745 4779 4414−812 −312 218 A C ATOM 1018 N PHE A 174 47.184 26.020 39.125 1.00 36.56A N ANISOU 1018 N PHE A 174 3674 5032 5186 −650 −339 802 A N ATOM 1019CA PHE A 174 46.785 26.850 37.999 1.00 47.05 A C ANISOU 1019 CA PHE A174 4869 6390 6615 −631 −369 1023 A C ATOM 1020 C PHE A 174 45.28326.750 37.899 1.00 60.55 A C ANISOU 1020 C PHE A 174 6506 8197 8302 −648−394 1067 A C ATOM 1021 O PHE A 174 44.771 25.655 37.957 1.00 57.31 A OANISOU 1021 O PHE A 174 6191 7935 7650 −726 −413 1035 A O ATOM 1022 CBPHE A 174 47.376 26.312 36.700 1.00 41.11 A C ANISOU 1022 CB PHE A 1744174 5766 5679 −671 −374 1179 A C ATOM 1023 CG PHE A 174 48.848 26.52036.567 1.00 42.57 A C ANISOU 1023 CG PHE A 174 4369 5883 5923 −639 −3421224 A C ATOM 1024 CD1 PHE A 174 49.357 27.772 36.312 1.00 42.26 A CANISOU 1024 CD1 PHE A 174 4184 5724 6147 −601 −368 1361 A C ATOM 1025CD2 PHE A 174 49.717 25.456 36.675 1.00 37.78 A C ANISOU 1025 CD2 PHE A174 3905 5329 5122 −646 −285 1157 A C ATOM 1026 CE1 PHE A 174 50.71227.964 36.182 1.00 42.70 A C ANISOU 1026 CE1 PHE A 174 4226 5738 6261−587 −350 1441 A C ATOM 1027 CE2 PHE A 174 51.070 25.639 36.547 1.0035.11 A C ANISOU 1027 CE2 PHE A 174 3540 4949 4850 −608 −246 1240 A CATOM 1028 CZ PHE A 174 51.571 26.895 36.301 1.00 37.24 A C ANISOU 1028CZ PHE A 174 3653 5123 5372 −588 −285 1388 A C ATOM 1029 N CYS A 17544.588 27.879 37.793 1.00 61.71 A N ANISOU 1029 N CYS A 175 6479 82558714 −578 −395 1160 A N ATOM 1030 CA CYS A 175 43.129 27.898 37.672 1.0066.01 A C ANISOU 1030 CA CYS A 175 6898 8901 9283 −581 −416 1270 A CATOM 1031 C CYS A 175 42.662 29.242 37.120 1.00 57.70 A C ANISOU 1031 CCYS A 175 5628 7749 8544 −500 −432 1494 A C ATOM 1032 O CYS A 175 43.44930.176 37.050 1.00 47.99 A O ANISOU 1032 O CYS A 175 4363 6347 7524 −442−424 1524 A O ATOM 1033 CB CYS A 175 42.466 27.637 39.013 1.00 63.25 A CANISOU 1033 CB CYS A 175 6557 8539 8936 −547 −352 1068 A C ATOM 1034 SGCYS A 175 42.297 29.145 39.959 1.00 61.36 A S ANISOU 1034 SG CYS A 1756225 8034 9055 −391 −248 930 A S ATOM 1035 N LYS A 176 41.393 29.34836.731 1.00 57.88 A N ANISOU 1035 N LYS A 176 5494 7882 8614 −505 −4641677 A N ATOM 1036 CA LYS A 176 40.874 30.605 36.183 1.00 55.23 A CANISOU 1036 CA LYS A 176 4926 7472 8586 −432 −490 1944 A C ATOM 1037 CLYS A 176 39.350 30.760 36.267 1.00 63.84 A C ANISOU 1037 C LYS A 1765838 8662 9757 −404 −484 2068 A C ATOM 1038 O LYS A 176 38.629 29.79136.085 1.00 73.90 A O ANISOU 1038 O LYS A 176 7174 10122 10783 −495 −5052014 A O ATOM 1039 CB LYS A 176 41.298 30.707 34.717 1.00 55.15 A CANISOU 1039 CB LYS A 176 4886 7588 8482 −538 −601 2224 A C ATOM 1040 CGLYS A 176 41.101 32.064 34.076 1.00 52.95 A C ANISOU 1040 CG LYS A 1764368 7203 8549 −472 −643 2525 A C ATOM 1041 CD LYS A 176 41.346 31.96132.585 1.00 58.00 A C ANISOU 1041 CD LYS A 176 4995 8028 9013 −611 −7552800 A C ATOM 1042 CE LYS A 176 41.197 33.301 31.891 1.00 72.41 A CANISOU 1042 CE LYS A 176 6594 9741 11179 −561 −809 3112 A C ATOM 1043 NZLYS A 176 41.327 33.159 30.416 1.00 71.50 A N1+ ANISOU 1043 NZ LYS A 1766474 9828 10863 −709 −909 3382 A N1+ ATOM 1044 N CYS A 177 38.862 31.96936.552 1.00 66.54 A N ANISOU 1044 N CYS A 177 5945 8874 10463 −277 −4582258 A N ATOM 1045 CA CYS A 177 37.421 32.210 36.574 1.00 73.32 A CANISOU 1045 CA CYS A 177 6575 9817 11464 −219 −437 2434 A C ATOM 1046 CCYS A 177 37.072 32.752 35.183 1.00 67.92 A C ANISOU 1046 C CYS A 1775637 9201 10967 −243 −549 2869 A C ATOM 1047 O CYS A 177 37.906 33.38534.541 1.00 70.67 A O ANISOU 1047 O CYS A 177 5967 9462 11421 −256 −6113008 A O ATOM 1048 CB CYS A 177 37.011 33.200 37.673 1.00 80.74 A CANISOU 1048 CB CYS A 177 7449 10517 12711 2 −251 2235 A C ATOM 1049 SGCYS A 177 37.404 32.751 39.387 1.00 90.22 A S ANISOU 1049 SG CYS A 1778896 11767 13615 −26 −148 1808 A S ATOM 1050 N PRO A 178 35.840 32.51234.711 1.00 63.67 A N ANISOU 1050 N PRO A 178 4885 8845 10461 −269 −5923124 A N ATOM 1051 CA PRO A 178 35.400 32.924 33.368 1.00 64.27 A CANISOU 1051 CA PRO A 178 4729 9060 10631 −360 −746 3583 A C ATOM 1052 CPRO A 178 35.217 34.427 33.128 1.00 68.37 A C ANISOU 1052 C PRO A 1785022 9316 11641 −172 −699 3780 A C ATOM 1053 O PRO A 178 35.038 35.19234.071 1.00 58.31 A O ANISOU 1053 O PRO A 178 3702 7757 10695 59 −5263603 A O ATOM 1054 CB PRO A 178 34.050 32.209 33.205 1.00 72.69 A CANISOU 1054 CB PRO A 178 5604 10364 11652 −419 −789 3793 A C ATOM 1055CG PRO A 178 33.968 31.224 34.325 1.00 70.63 A C ANISOU 1055 CG PRO A178 5568 10163 11105 −446 −705 3418 A C ATOM 1056 CD PRO A 178 34.77831.805 35.437 1.00 57.85 A C ANISOU 1056 CD PRO A 178 4124 8250 9606−270 −533 3030 A C ATOM 1057 N SER A 179 35.252 34.820 31.852 1.00 75.00A N ANISOU 1057 N SER A 179 5736 10246 12514 −286 −856 4144 A N ATOM1058 CA SER A 179 35.072 36.212 31.420 1.00 77.18 A C ANISOU 1058 CA SERA 179 5771 10303 13251 −146 −849 4429 A C ATOM 1059 C SER A 179 33.58636.536 31.338 1.00 84.91 A C ANISOU 1059 C SER A 179 6558 11293 14413−77 −800 4676 A C ATOM 1060 O SER A 179 32.778 35.603 31.394 1.00 93.34A O ANISOU 1060 O SER A 179 7611 12595 15260 −160 −821 4702 A O ATOM1061 CB SER A 179 35.708 36.422 30.052 1.00 75.06 A C ANISOU 1061 CB SERA 179 5547 10127 12846 −332 −1006 4701 A C ATOM 1062 OG SER A 179 35.15135.532 29.103 1.00 72.12 A O ANISOU 1062 OG SER A 179 5163 10066 12175−557 −1143 4993 A O ATOM 1063 N LYS A 180 33.242 37.844 31.319 1.0085.37 A N ANISOU 1063 N LYS A 180 6460 11089 14888 67 −735 4876 A N ATOM1064 CA LYS A 180 31.842 38.253 31.252 1.00 93.10 A C ANISOU 1064 CA LYSA 180 7231 12071 16073 136 −675 5147 A C ATOM 1065 C LYS A 180 31.11537.485 30.153 1.00 80.47 A C ANISOU 1065 C LYS A 180 5617 10844 14113−148 −863 5486 A C ATOM 1066 O LYS A 180 30.079 36.848 30.401 1.00 73.21A O ANISOU 1066 O LYS A 180 4637 10107 13075 −178 −856 5549 A O ATOM1067 CB LYS A 180 31.741 39.761 31.025 1.00 105.68 A C ANISOU 1067 CBLYS A 180 8666 13335 18155 284 −598 5350 A C ATOM 1068 CG LYS A 18030.329 40.304 31.192 1.00 111.80 A C ANISOU 1068 CG LYS A 180 9205 1404919225 418 −479 5588 A C ATOM 1069 CD LYS A 180 30.255 41.820 30.983 1.00115.29 A C ANISOU 1069 CD LYS A 180 9487 14143 20175 560 −393 5777 A CATOM 1070 CE LYS A 180 30.854 42.596 32.153 1.00 114.37 A C ANISOU 1070CE LYS A 180 9436 13608 20413 852 −187 5389 A C ATOM 1071 NZ LYS A 18030.531 44.055 32.078 1.00 116.90 A N1+ ANISOU 1071 NZ LYS A 180 959013575 21254 1011 −74 5559 A N1+ ATOM 1072 N ASN A 181 31.691 37.49528.941 1.00 73.66 A N ANISOU 1072 N ASN A 181 4827 10108 13052 −374−1033 5698 A N ATOM 1073 CA ASN A 181 31.139 36.760 27.807 1.00 75.90 AC ANISOU 1073 CA ASN A 181 5137 10746 12957 −680 −1221 6005 A C ATOM1074 C ASN A 181 30.844 35.302 28.159 1.00 72.41 A C ANISOU 1074 C ASN A181 4842 10580 12092 −812 −1280 5813 A C ATOM 1075 O ASN A 181 29.76034.789 27.869 1.00 75.78 A O ANISOU 1075 O ASN A 181 5209 11215 12370−950 −1354 6032 A O ATOM 1076 CB ASN A 181 32.101 36.850 26.618 1.0082.18 A C ANISOU 1076 CB ASN A 181 6038 11644 13541 −894 −1360 6143 A CATOM 1077 CG ASN A 181 32.109 38.228 25.981 1.00 102.84 A C ANISOU 1077CG ASN A 181 8473 14064 16536 −857 −1346 6463 A C ATOM 1078 OD1 ASN A181 31.496 39.165 26.500 1.00 112.69 A O ANISOU 1078 OD1 ASN A 181 952915052 18234 −648 −1221 6543 A O ATOM 1079 ND2 ASN A 181 32.801 38.36124.855 1.00 106.55 A N ANISOU 1079 ND2 ASN A 181 8993 14661 16830 −1069−1461 6647 A N ATOM 1080 N CBN A 182 31.792 34.613 28.785 1.00 74.11 A NANISOU 1080 N CBN A 182 5245 10797 12115 −789 −1252 5418 A N ATOM 1081CA CBN A 182 31.560 33.205 29.089 1.00 82.34 A C ANISOU 1081 CA CBN A182 6437 12093 12756 −944 −1314 5236 A C ATOM 1082 C CBN A 182 30.46433.033 30.130 1.00 94.59 A C ANISOU 1082 C CBN A 182 7857 13622 14460−798 −1197 5178 A C ATOM 1083 O CBN A 182 29.760 32.017 30.125 1.00108.30 A O ANISOU 1083 O CBN A 182 9647 15596 15904 −969 −1281 5210 A OATOM 1084 CB CBN A 182 32.848 32.543 29.568 1.00 76.61 A C ANISOU 1084CB CBN A 182 5924 11360 11824 −954 −1295 4838 A C ATOM 1085 CG CBN A 18233.959 32.506 28.529 1.00 68.61 A C ANISOU 1085 CG CBN A 182 5055 1042810587 −1118 −1407 4887 A C ATOM 1086 CD CBN A 182 35.307 32.146 29.1431.00 65.51 A C ANISOU 1086 CD CBN A 182 4881 9921 10089 −1052 −1318 4479A C ATOM 1087 OE1 CBN A 182 35.849 32.884 29.976 1.00 70.21 A O ANISOU1087 OE1 CBN A 182 5467 10218 10993 −821 −1165 4278 A O ATOM 1088 NE2CBN A 182 35.856 31.005 28.740 1.00 56.66 A N ANISOU 1088 NE2 CBN A 1824065 8976 8487 −1252 −1373 4307 A N ATOM 1089 N BEU A 183 30.293 34.00231.032 1.00 91.16 A N ANISOU 1089 N BEU A 183 7257 12906 14474 −491 −9965093 A N ATOM 1090 CA BEU A 183 29.198 33.892 31.994 1.00 85.89 A CANISOU 1090 CA BEU A 183 6443 12236 13954 −338 −857 5062 A C ATOM 1091 CBEU A 183 27.841 34.054 31.297 1.00 85.84 A C ANISOU 1091 C BEU A 1836258 12368 13990 −429 −932 5515 A C ATOM 1092 O BEU A 183 26.934 33.24031.499 1.00 86.51 A O ANISOU 1092 O BEU A 183 6319 12666 13886 −529 −9715588 A O ATOM 1093 CB BEU A 183 29.388 34.901 33.136 1.00 83.75 A CANISOU 1093 CB BEU A 183 6061 11619 14140 18 −597 4832 A C ATOM 1094 CGBEU A 183 30.509 34.593 34.150 1.00 69.40 A C ANISOU 1094 CG BEU A 1834399 9685 12284 115 −491 4363 A C ATOM 1095 CD1 BEU A 183 30.735 35.71735.176 1.00 69.50 A C ANISOU 1095 CD1 BEU A 183 4329 9316 12761 456 −2384147 A C ATOM 1096 CD2 BEU A 183 30.211 33.299 34.877 1.00 62.96 A CANISOU 1096 CD2 BEU A 183 3661 9112 11148 17 −482 4158 A C ATOM 1097 NASN A 184 27.707 35.056 30.410 1.00 78.48 A N ANISOU 1097 N ASN A 1845202 11331 13286 −432 −971 5851 A N ATOM 1098 CA ASN A 184 26.456 35.22229.662 1.00 77.04 A C ANISOU 1098 CA ASN A 184 4838 11289 13144 −552−1055 6327 A C ATOM 1099 C ASN A 184 26.118 33.978 28.838 1.00 76.55 A CANISOU 1099 C ASN A 184 4917 11598 12571 −924 −1292 6487 A C ATOM 1100 OASN A 184 24.959 33.562 28.797 1.00 80.77 A O ANISOU 1100 O ASN A 1845347 12304 13039 −1018 −1338 6730 A O ATOM 1101 CB ASN A 184 26.51636.466 28.766 1.00 87.69 A C ANISOU 1101 CB ASN A 184 6044 12475 14798−541 −1071 6665 A C ATOM 1102 CG ASN A 184 25.140 36.836 28.159 1.00107.46 A C ANISOU 1102 CG ASN A 184 8300 15076 17454 −619 −1111 7183 A CATOM 1103 OD1 ASN A 184 24.950 36.785 26.943 1.00 116.57 A O ANISOU 1103OD1 ASN A 184 9442 16411 18438 −899 −1294 7548 A O ATOM 1104 ND2 ASN A184 24.189 37.229 29.013 1.00 107.94 A N ANISOU 1104 ND2 ASN A 184 815415019 17839 −373 −926 7221 A N ATOM 1105 N BYS A 185 27.111 33.36428.182 1.00 79.07 A N ANISOU 1105 N BYS A 185 5479 12041 12524 −1147−1438 6356 A N ATOM 1106 CA BYS A 185 26.912 32.079 27.494 1.00 80.45 AC ANISOU 1106 CA BYS A 185 5842 12545 12182 −1512 −1641 6417 A C ATOM1107 C BYS A 185 26.541 30.944 28.440 1.00 71.90 A C ANISOU 1107 C BYS A185 4855 11574 10890 −1525 −1624 6158 A C ATOM 1108 O BYS A 185 26.22429.843 27.979 1.00 71.82 A O ANISOU 1108 O BYS A 185 5002 11810 10476−1834 −1782 6207 A O ATOM 1109 CB LYS A 185 28.174 31.616 26.742 1.0085.85 A C ANISOU 1109 CB LYS A 185 6783 13317 12517 −1719 −1752 6249 A CATOM 1110 CG LYS A 185 28.488 32.285 25.405 1.00 92.80 A C ANISOU 1110CG LYS A 185 7626 14242 13391 −1890 −1843 6563 A C ATOM 1111 CD LYS A185 29.791 31.714 24.839 1.00 77.13 A C ANISOU 1111 CD LYS A 185 589612362 11048 −2070 −1910 6327 A C ATOM 1112 CE LYS A 185 30.209 32.41523.566 1.00 72.78 A C ANISOU 1112 CE LYS A 185 5289 11863 10500 −2206−1978 6537 A C ATOM 1113 NZ LYS A 185 31.489 31.872 23.067 1.00 79.48 AN1+ ANISOU 1113 NZ LYS A 185 6357 12821 11019 −2346 −2018 6265 A N1+ATOM 1114 N GLY A 186 26.624 31.154 29.742 1.00 70.63 A N ANISOU 1114 NGLY A 186 4621 11238 10979 −1227 −1429 5869 A N ATOM 1115 CA GLY A 18626.231 30.118 30.660 1.00 70.28 A C ANISOU 1115 CA GLY A 186 4643 1131110750 −1249 −1403 5651 A C ATOM 1116 C GLY A 186 27.341 29.201 31.0961.00 65.52 A C ANISOU 1116 C GLY A 186 4307 10733 9854 −1325 −1418 5219A C ATOM 1117 O GLY A 186 27.051 28.125 31.640 1.00 64.43 A O ANISOU1117 O GLY A 186 4278 10735 9469 −1439 −1450 5067 A O ATOM 1118 N ILE A187 28.597 29.588 30.867 1.00 65.47 A N ANISOU 1118 N ILE A 187 440810594 9872 −1276 −1400 5037 A N ATOM 1119 CA ILE A 187 29.761 28.85531.363 1.00 73.01 A C ANISOU 1119 CA ILE A 187 5591 11539 10611 −1312−1384 4629 A C ATOM 1120 C ILE A 187 29.946 29.228 32.831 1.00 77.96 A CANISOU 1120 C ILE A 187 6111 11970 11540 −1014 −1150 4333 A C ATOM 1121O ILE A 187 30.180 30.397 33.160 1.00 84.30 A O ANISOU 1121 O ILE A 1876762 12531 12738 −754 −999 4322 A O ATOM 1122 CB ILE A 187 31.027 29.18930.545 1.00 69.03 A C ANISOU 1122 CB ILE A 187 5215 10976 10036 −1367−1443 4589 A C ATOM 1123 CG1 ILE A 187 30.790 29.057 29.028 1.00 61.27 AC ANISOU 1123 CG1 ILE A 187 4300 10185 8794 −1648 −1641 4923 A C ATOM1124 CG2 ILE A 187 32.223 28.355 30.985 1.00 58.51 A C ANISOU 1124 CG2ILE A 187 4185 9612 8433 −1412 −1408 4167 A C ATOM 1125 CD1 ILE A 18730.728 27.630 28.502 1.00 60.46 A C ANISOU 1125 CD1 ILE A 187 4467 103388170 −1995 −1807 4861 A C ATOM 1126 N LYS A 188 29.814 28.248 33.7221.00 72.39 A N ANISOU 1126 N LYS A 188 5492 11363 10651 −1064 −1117 4097A N ATOM 1127 CA LYS A 188 30.026 28.530 35.133 1.00 72.37 A C ANISOU1127 CA LYS A 188 5504 11153 10839 −797 −864 3756 A C ATOM 1128 C LYS A188 31.358 28.002 35.655 1.00 62.02 A C ANISOU 1128 C LYS A 188 45849693 9289 −802 −799 3292 A C ATOM 1129 O LYS A 188 31.783 28.403 36.7441.00 54.58 A O ANISOU 1129 O LYS A 188 3698 8541 8500 −592 −599 2991 A OATOM 1130 CB LYS A 188 28.860 27.974 35.962 1.00 77.56 A C ANISOU 1130CB LYS A 188 6003 11977 11489 −789 −809 3804 A C ATOM 1131 CG LYS A 18827.510 28.520 35.480 1.00 89.47 A C ANISOU 1131 CG LYS A 188 7242 1355913195 −747 −835 4227 A C ATOM 1132 CD LYS A 188 26.347 28.191 36.3961.00 92.88 A C ANISOU 1132 CD LYS A 188 7521 14101 13669 −669 −721 4267A C ATOM 1133 CE LYS A 188 26.032 26.702 36.372 1.00 94.49 A C ANISOU1133 CE LYS A 188 7902 14562 13436 −975 −902 4250 A C ATOM 1134 NZ LYS A188 25.328 26.308 35.117 1.00 99.84 A N1+ ANISOU 1134 NZ LYS A 188 862415407 13905 −1245 −1152 4624 A N1+ ATOM 1135 N TYR A 189 32.053 27.16134.893 1.00 60.77 A N ANISOU 1135 N TYR A 189 4695 9628 8768 −1034 −9543235 A N ATOM 1136 CA TYR A 189 33.314 26.634 35.384 1.00 54.85 A CANISOU 1136 CA TYR A 189 4283 8740 7817 −1025 −884 2833 A C ATOM 1137 CTYR A 189 34.344 26.543 34.269 1.00 53.00 A C ANISOU 1137 C TYR A 1894243 8499 7396 −1147 −986 2851 A C ATOM 1138 O TYR A 189 34.051 26.01833.195 1.00 51.29 A O ANISOU 1138 O TYR A 189 4068 8480 6940 −1372 −11583061 A O ATOM 1139 CB TYR A 189 33.092 25.260 36.015 1.00 47.86 A CANISOU 1139 CB TYR A 189 3588 7981 6615 −1168 −915 2642 A C ATOM 1140 CGTYR A 189 32.078 25.277 37.146 1.00 48.77 A C ANISOU 1140 CG TYR A 1893510 8142 6879 −1058 −804 2631 A C ATOM 1141 CD1 TYR A 189 32.314 25.99938.307 1.00 52.31 A C ANISOU 1141 CD1 TYR A 189 3908 8396 7572 −805 −5832402 A C ATOM 1142 CD2 TYR A 189 30.903 24.558 37.057 1.00 52.35 A CANISOU 1142 CD2 TYR A 189 3841 8840 7210 −1223 −919 2849 A C ATOM 1143CE1 TYR A 189 31.398 26.015 39.331 1.00 64.50 A C ANISOU 1143 CE1 TYR A189 5284 9997 9227 −697 −453 2383 A C ATOM 1144 CE2 TYR A 189 29.96924.567 38.083 1.00 63.04 A C ANISOU 1144 CE2 TYR A 189 4995 10263 8695−1117 −801 2864 A C ATOM 1145 CZ TYR A 189 30.218 25.298 39.220 1.0065.24 A C ANISOU 1145 CZ TYR A 189 5226 10354 9208 −843 −554 2624 A CATOM 1146 OH TYR A 189 29.297 25.297 40.246 1.00 62.96 A O ANISOU 1146OH TYR A 189 4748 10150 9024 −732 −409 2627 A O ATOM 1147 N LEU A 19035.577 26.917 34.598 1.00 52.03 A N ANISOU 1147 N LEU A 190 4237 81597371 −1009 −878 2639 A N ATOM 1148 CA LEU A 190 36.736 26.753 33.7301.00 49.76 A C ANISOU 1148 CA LEU A 190 4184 7867 6857 −1107 −927 2573 AC ATOM 1149 C LEU A 190 37.484 25.706 34.553 1.00 50.35 A C ANISOU 1149C LEU A 190 4550 7880 6699 −1118 −856 2206 A C ATOM 1150 O LEU A 19038.005 26.013 35.619 1.00 59.49 A O ANISOU 1150 O LEU A 190 5744 88518010 −962 −727 1973 A O ATOM 1151 CB LEU A 190 37.537 28.040 33.617 1.0049.86 A C ANISOU 1151 CB LEU A 190 4107 7697 7141 −966 −872 2636 A CATOM 1152 CG LEU A 190 36.985 29.101 32.669 1.00 48.62 A C ANISOU 1152CG LEU A 190 3647 7542 7283 −922 −931 3007 A C ATOM 1153 CD1 LEU A 19037.837 30.357 32.717 1.00 48.76 A C ANISOU 1153 CD1 LEU A 190 3632 73687526 −813 −891 3042 A C ATOM 1154 CD2 LEU A 190 36.923 28.549 31.2591.00 59.49 A C ANISOU 1154 CD2 LEU A 190 5001 9200 8405 −1161 −1112 3307A C ATOM 1155 N ILE A 191 37.534 24.474 34.059 1.00 43.06 A N ANISOU1155 N ILE A 191 3843 7106 5412 −1313 −947 2164 A N ATOM 1156 CA ILE A191 38.080 23.350 34.816 1.00 41.24 A C ANISOU 1156 CA ILE A 191 38706834 4967 −1341 −898 1860 A C ATOM 1157 C ILE A 191 39.513 23.103 34.3581.00 44.18 A C ANISOU 1157 C ILE A 191 4467 7119 5201 −1328 −849 1724 AC ATOM 1158 O ILE A 191 39.759 22.857 33.178 1.00 63.43 A O ANISOU 1158O ILE A 191 7008 9653 7441 −1445 −913 1839 A O ATOM 1159 CB ILE A 19137.213 22.092 34.634 1.00 41.71 A C ANISOU 1159 CB ILE A 191 4045 70734731 −1560 −1024 1893 A C ATOM 1160 CG1 ILE A 191 35.798 22.304 35.2011.00 48.59 A C ANISOU 1160 CG1 ILE A 191 4660 8050 5753 −1564 −1060 2049A C ATOM 1161 CG2 ILE A 191 37.880 20.886 35.240 1.00 40.88 A C ANISOU1161 CG2 ILE A 191 4225 6905 4404 −1600 −987 1606 A C ATOM 1162 CD1 ILEA 191 35.040 21.017 35.506 1.00 43.05 A C ANISOU 1162 CD1 ILE A 191 40677487 4801 −1756 −1162 2022 A C ATOM 1163 N THR A 192 40.453 23.13835.279 1.00 54.21 A N ANISOU 1163 N THR A 192 5810 8222 6564 −1194 −7321493 A N ATOM 1164 CA THR A 192 41.850 22.964 34.928 1.00 37.84 A CANISOU 1164 CA THR A 192 3898 6069 4409 −1158 −671 1406 A C ATOM 1165 CTHR A 192 42.112 21.488 34.716 1.00 37.19 A C ANISOU 1165 C THR A 1924098 6030 4001 −1265 −675 1251 A C ATOM 1166 O THR A 192 41.721 20.66435.517 1.00 36.39 A O ANISOU 1166 O THR A 192 4074 5913 3841 −1289 −6751098 A O ATOM 1167 CB THR A 192 42.770 23.641 35.946 1.00 36.84 A CANISOU 1167 CB THR A 192 3714 5750 4534 −991 −567 1267 A C ATOM 1168 OG1THR A 192 42.544 23.095 37.239 1.00 37.49 A O ANISOU 1168 OG1 THR A 1923994 5771 4480 −984 −508 1056 A O ATOM 1169 CG2 THR A 192 42.518 25.14735.952 1.00 37.08 A C ANISOU 1169 CG2 THR A 192 3572 5717 4801 −911 −5431227 A C ATOM 1170 N TYR A 193 42.805 21.173 33.631 1.00 37.82 A NANISOU 1170 N TYR A 193 4334 6167 3870 −1329 −673 1301 A N ATOM 1171 CATYR A 193 42.999 19.788 33.209 1.00 38.00 A C ANISOU 1171 CA TYR A 1934651 6229 3560 −1446 −677 1182 A C ATOM 1172 C TYR A 193 44.421 19.60332.695 1.00 46.25 A C ANISOU 1172 C TYR A 193 5845 7206 4524 −1357 −5461113 A C ATOM 1173 O TYR A 193 44.898 20.407 31.893 1.00 49.56 A OANISOU 1173 O TYR A 193 6175 7664 4992 −1315 −515 1259 A O ATOM 1174 CBTYR A 193 42.000 19.422 32.106 1.00 39.82 A C ANISOU 1174 CB TYR A 1934949 6637 3544 −1660 −814 1338 A C ATOM 1175 CG TYR A 193 42.226 18.07131.470 1.00 40.61 A C ANISOU 1175 CG TYR A 193 5397 6757 3277 −1799 −8171210 A C ATOM 1176 CD1 TYR A 193 41.845 16.911 32.123 1.00 40.22 A CANISOU 1176 CD1 TYR A 193 5519 6659 3105 −1883 −859 1051 A C ATOM 1177CD2 TYR A 193 42.822 17.948 30.219 1.00 42.03 A C ANISOU 1177 CD2 TYR A193 5745 6996 3228 −1847 −771 1246 A C ATOM 1178 CE1 TYR A 193 42.05515.664 31.546 1.00 42.08 A C ANISOU 1178 CE1 TYR A 193 6101 6868 3021−2008 −858 919 A C ATOM 1179 CE2 TYR A 193 43.054 16.698 29.650 1.0043.13 A C ANISOU 1179 CE2 TYR A 193 6241 7122 3026 −1961 −745 1093 A CATOM 1180 CZ TYR A 193 42.658 15.563 30.319 1.00 42.78 A C ANISOU 1180CZ TYR A 193 6375 6993 2886 −2039 −793 925 A C ATOM 1181 OH TYR A 19342.873 14.306 29.797 1.00 44.14 A O ANISOU 1181 OH TYR A 193 6923 71062743 −2149 −766 758 A O ATOM 1182 N VAL A 194 45.115 18.559 33.141 1.0051.86 A N ANISOU 1182 N VAL A 194 6761 7819 5123 −1321 −464 917 A N ATOM1183 CA VAL A 194 46.489 18.347 32.692 1.00 45.92 A C ANISOU 1183 CA VALA 194 6127 7004 4317 −1210 −313 870 A C ATOM 1184 C VAL A 194 46.46617.418 31.483 1.00 39.33 A C ANISOU 1184 C VAL A 194 5572 6244 3128−1322 −288 843 A C ATOM 1185 O VAL A 194 46.066 16.253 31.582 1.00 39.54A O ANISOU 1185 O VAL A 194 5828 6237 2958 −1422 −319 706 A O ATOM 1186CB VAL A 194 47.379 17.803 33.817 1.00 44.75 A C ANISOU 1186 CB VAL A194 6023 6701 4281 −1086 −221 706 A C ATOM 1187 CG1 VAL A 194 48.58517.068 33.240 1.00 40.42 A C ANISOU 1187 CG1 VAL A 194 5664 6097 3596−997 −59 647 A C ATOM 1188 CG2 VAL A 194 47.871 18.956 34.684 1.00 44.46A C ANISOU 1188 CG2 VAL A 194 5732 6595 4567 −966 −209 755 A C ATOM 1189N TRP A 195 46.906 17.945 30.344 1.00 45.74 A N ANISOU 1189 N TRP A 1956375 7153 3853 −1315 −232 974 A N ATOM 1190 CA TRP A 195 46.798 17.27129.062 1.00 42.69 A C ANISOU 1190 CA TRP A 195 6250 6871 3098 −1446 −211966 A C ATOM 1191 C TRP A 195 47.727 16.072 28.989 1.00 52.83 A C ANISOU1191 C TRP A 195 7833 8033 4207 −1360 −30 753 A C ATOM 1192 O TRP A 19548.924 16.181 29.264 1.00 61.98 A O ANISOU 1192 O TRP A 195 8938 91035510 −1159 141 728 A O ATOM 1193 CB TRP A 195 47.116 18.270 27.959 1.0044.02 A C ANISOU 1193 CB TRP A 195 6292 7191 3242 −1447 −185 1183 A CATOM 1194 CG TRP A 195 46.815 17.824 26.545 1.00 55.90 A C ANISOU 1194CG TRP A 195 8034 8862 4344 −1631 −198 1222 A C ATOM 1195 CD1 TRP A 19547.702 17.288 25.662 1.00 59.43 A C ANISOU 1195 CD1 TRP A 195 8721 93364523 −1591 −11 1141 A C ATOM 1196 CD2 TRP A 195 45.561 17.929 25.8381.00 59.45 A C ANISOU 1196 CD2 TRP A 195 8496 9488 4605 −1891 −404 1368A C ATOM 1197 NE1 TRP A 195 47.086 17.033 24.463 1.00 68.01 A N ANISOU1197 NE1 TRP A 195 10003 10601 5238 −1822 −87 1196 A N ATOM 1198 CE2 TRPA 195 45.773 17.418 24.537 1.00 64.48 A C ANISOU 1198 CE2 TRP A 195 941510254 4832 −2023 −344 1350 A C ATOM 1199 CE3 TRP A 195 44.284 18.38926.180 1.00 65.27 A C ANISOU 1199 CE3 TRP A 195 9029 10294 5477 −2025−626 1523 A C ATOM 1200 CZ2 TRP A 195 44.757 17.358 23.571 1.00 53.06 AC ANISOU 1200 CZ2 TRP A 195 8043 8985 3132 −2296 −524 1471 A C ATOM 1201CZ3 TRP A 195 43.262 18.325 25.208 1.00 66.43 A C ANISOU 1201 CZ3 TRP A195 9235 10644 5360 −2303 −807 1690 A C ATOM 1202 CH2 TRP A 195 43.51617.817 23.923 1.00 60.06 A C ANISOU 1202 CH2 TRP A 195 8692 9925 4202−2424 −759 1641 A C ATOM 1203 N GLN A 196 47.170 14.928 28.614 1.0047.18 A N ANISOU 1203 N GLN A 196 7429 7303 3194 −1516 −70 614 A N ATOM1204 CA GLN A 196 47.888 13.665 28.606 1.00 56.27 A C ANISOU 1204 CA GLNA 196 8896 8294 4192 −1439 95 392 A C ATOM 1205 C GLN A 196 48.45413.347 27.225 1.00 74.85 A C ANISOU 1205 C GLN A 196 11508 10711 6221−1446 261 354 A C ATOM 1206 O GLN A 196 48.047 13.913 26.202 1.00 69.83A O ANISOU 1206 O GLN A 196 10863 10271 5399 −1588 197 491 A O ATOM 1207CB GLN A 196 46.976 12.513 29.044 1.00 55.67 A C ANISOU 1207 CB GLN A196 9048 8122 3980 −1612 −41 240 A C ATOM 1208 CG GLN A 196 46.53112.514 30.503 1.00 43.32 A C ANISOU 1208 CG GLN A 196 7288 6479 2694−1591 −160 233 A C ATOM 1209 CD GLN A 196 47.678 12.692 31.466 1.0049.26 A C ANISOU 1209 CD GLN A 196 7883 7097 3738 −1337 −10 199 A C ATOM1210 OE1 GLN A 196 48.449 11.766 31.713 1.00 57.39 A O ANISOU 1210 OE1GLN A 196 9089 7961 4757 −1228 126 63 A O ATOM 1211 NE2 GLN A 196 47.79613.890 32.025 1.00 53.64 A N ANISOU 1211 NE2 GLN A 196 8104 7713 4564−1251 −41 335 A N ATOM 1212 N ASP A 197 49.405 12.409 27.220 1.00 91.15A N ANISOU 1212 N ASP A 197 13805 12607 8220 −1288 486 172 A N ATOM 1213CA ASP A 197 49.941 11.846 25.988 1.00 98.99 A C ANISOU 1213 CA ASP A197 15119 13621 8871 −1277 689 70 A C ATOM 1214 C ASP A 197 48.80211.261 25.169 1.00 90.90 A C ANISOU 1214 C ASP A 197 14415 12672 7453−1594 525 −11 A C ATOM 1215 O ASP A 197 48.024 10.432 25.660 1.00 89.37A O ANISOU 1215 O ASP A 197 14390 12357 7211 −1743 373 −134 A O ATOM1216 CB ASP A 197 50.994 10.773 26.315 1.00 107.68 A C ANISOU 1216 CBASP A 197 16426 14476 10011 −1046 952 −130 A C ATOM 1217 CG ASP A 19751.927 10.465 25.138 1.00 111.71 A C ANISOU 1217 CG ASP A 197 1717215012 10262 −919 1260 −203 A C ATOM 1218 OD1 ASP A 197 51.861 11.16424.100 1.00 114.07 A O ANISOU 1218 OD1 ASP A 197 17447 15542 10354 −10071273 −83 A O ATOM 1219 OD2 ASP A 197 52.732 9.514 25.268 1.00 111.30 AO1− ANISOU 1219 OD2 ASP A 197 17321 14748 10219 −723 1501 −371 A O1−ATOM 1220 N ASN A 198 48.661 11.749 23.941 1.00 94.34 A N ANISOU 1220 NASN A 198 14894 13319 7630 −1715 525 91 A N ATOM 1221 CA ASN A 19847.660 11.256 23.000 1.00 100.27 A C ANISOU 1221 CA ASN A 198 1580314110 8184 −1945 348 40 A C ATOM 1222 C ASN A 198 46.232 11.537 23.4751.00 87.84 A C ANISOU 1222 C ASN A 198 14031 12593 6754 −2174 8 176 A CATOM 1223 O ASN A 198 45.317 10.764 23.185 1.00 97.24 A O ANISOU 1223 OASN A 198 15376 13719 7850 −2370 −149 95 A O ATOM 1224 CB ASN A 19847.843 9.753 22.703 1.00 108.71 A C ANISOU 1224 CB ASN A 198 17283 149439078 −1933 466 −265 A C ATOM 1225 CG ASN A 198 49.178 9.425 21.981 1.00105.86 A C ANISOU 1225 CG ASN A 198 17131 14541 8550 −1703 831 −402 A CATOM 1226 OD1 ASN A 198 49.709 10.232 21.206 1.00 100.80 A O ANISOU 1226OD1 ASN A 198 16376 14108 7815 −1639 951 −264 A O ATOM 1227 ND2 ASN A198 49.707 8.224 22.238 1.00 103.79 A N ANISOU 1227 ND2 ASN A 198 1716014004 8270 −1573 1012 −657 A N ATOM 1228 N ASP A 199 46.023 12.62424.219 1.00 66.96 A N ANISOU 1228 N ASP A 199 11035 10061 4346 −2151 −97393 A N ATOM 1229 CA ASP A 199 44.699 13.227 24.319 1.00 59.25 A CANISOU 1229 CA ASP A 199 9804 9212 3496 −2349 −384 597 A C ATOM 1230 CASP A 199 44.310 13.903 22.992 1.00 70.77 A C ANISOU 1230 C ASP A 19911185 10881 4823 −2492 −462 784 A C ATOM 1231 O ASP A 199 45.166 14.36822.235 1.00 59.16 A O ANISOU 1231 O ASP A 199 9728 9508 3241 −2397 −302835 A O ATOM 1232 CB ASP A 199 44.672 14.255 25.452 1.00 58.12 A CANISOU 1232 CB ASP A 199 9309 9126 3649 −2257 −444 777 A C ATOM 1233 CGASP A 199 44.193 13.672 26.788 1.00 75.22 A C ANISOU 1233 CG ASP A 19911460 11147 5972 −2258 −534 669 A C ATOM 1234 OD1 ASP A 199 43.88612.465 26.837 1.00 82.68 A O ANISOU 1234 OD1 ASP A 199 12655 11940 6822−2331 −556 475 A O ATOM 1235 OD2 ASP A 199 44.121 14.425 27.793 1.0068.22 A O1− ANISOU 1235 OD2 ASP A 199 10245 10238 5439 −2126 −564 761 AO1− ATOM 1236 N ASN A 200 43.001 13.915 22.690 1.00 73.30 A N ANISOU1236 N ASN A 200 11420 11276 5153 −2732 −704 901 A N ATOM 1237 CA ASN A200 42.438 14.644 21.553 1.00 74.06 A C ANISOU 1237 CA ASN A 200 1138311583 5173 −2905 −827 1128 A C ATOM 1238 C ASN A 200 41.486 15.71822.053 1.00 66.84 A C ANISOU 1238 C ASN A 200 10053 10794 4548 −2973−1031 1429 A C ATOM 1239 O ASN A 200 40.981 15.651 23.178 1.00 65.28 A OANISOU 1239 O ASN A 200 9725 10522 4558 −2945 −1109 1431 A O ATOM 1240CB ASN A 200 41.635 13.755 20.587 1.00 67.94 A C ANISOU 1240 CB ASN A200 10855 10810 4150 −3174 −943 1052 A C ATOM 1241 CG ASN A 200 42.47112.708 19.924 1.00 66.46 A C ANISOU 1241 CG ASN A 200 11090 10504 3659−3124 −745 765 A C ATOM 1242 OD1 ASN A 200 42.072 11.552 19.834 1.0067.77 A O ANISOU 1242 OD1 ASN A 200 11535 10522 3691 −3252 −783 578 A OATOM 1243 ND2 ASN A 200 43.648 13.096 19.467 1.00 66.40 A N ANISOU 1243ND2 ASN A 200 11126 10553 3549 −2932 −520 739 A N ATOM 1244 N VAL A 20141.194 16.680 21.181 1.00 68.94 A N ANISOU 1244 N VAL A 201 6377 152334583 −2088 787 548 A N ATOM 1245 CA VAL A 201 40.238 17.711 21.546 1.0067.74 A C ANISOU 1245 CA VAL A 201 6240 15038 4458 −2096 678 889 A CATOM 1246 C VAL A 201 38.830 17.136 21.602 1.00 67.39 A C ANISOU 1246 CVAL A 201 6265 15073 4265 −2155 531 715 A C ATOM 1247 O VAL A 201 38.03817.456 22.500 1.00 67.52 A O ANISOU 1247 O VAL A 201 6362 14834 4456−2173 398 818 A O ATOM 1248 CB VAL A 201 40.334 18.891 20.569 1.00 69.85A C ANISOU 1248 CB VAL A 201 6402 15540 4597 −2030 731 1244 A C ATOM1249 CG1 VAL A 201 39.276 19.921 20.921 1.00 68.77 A C ANISOU 1249 CG1VAL A 201 6297 15277 4554 −1998 583 1552 A C ATOM 1250 CG2 VAL A 20141.753 19.478 20.621 1.00 70.14 A C ANISOU 1250 CG2 VAL A 201 6374 154614814 −1994 897 1400 A C ATOM 1251 N THR A 202 38.489 16.281 20.647 1.0069.62 A N ANISOU 1251 N THR A 202 6509 15687 4256 −2182 550 429 A N ATOM1252 CA THR A 202 37.164 15.671 20.663 1.00 69.57 A C ANISOU 1252 CA THRA 202 6547 15721 4165 −2239 417 207 A C ATOM 1253 C THR A 202 36.97814.797 21.895 1.00 67.08 A C ANISOU 1253 C THR A 202 6396 15004 4088−2305 366 −62 A C ATOM 1254 O THR A 202 35.919 14.816 22.530 1.00 71.06A O ANISOU 1254 O THR A 202 6974 15360 4667 −2354 241 −70 A O ATOM 1255CB THR A 202 36.952 14.836 19.406 1.00 72.63 A C ANISOU 1255 CB THR A202 6843 16436 4319 −2237 448 −95 A C ATOM 1256 OG1 THR A 202 37.36215.594 18.266 1.00 78.79 A O ANISOU 1256 OG1 THR A 202 7490 17493 4954−2141 502 142 A O ATOM 1257 CG2 THR A 202 35.500 14.439 19.286 1.0073.02 A C ANISOU 1257 CG2 THR A 202 6885 16550 4308 −2283 325 −270 A CATOM 1258 N LEU A 203 38.001 14.018 22.241 1.00 66.72 A N ANISOU 1258 NLEU A 203 6413 14711 4226 −2280 451 −291 A N ATOM 1259 CA LEU A 20337.918 13.096 23.367 1.00 68.17 A C ANISOU 1259 CA LEU A 203 6775 144344694 −2298 410 −562 A C ATOM 1260 C LEU A 203 37.710 13.851 24.678 1.0067.47 A C ANISOU 1260 C LEU A 203 6781 13919 4935 −2267 302 −299 A CATOM 1261 O LEU A 203 36.740 13.613 25.417 1.00 60.16 A O ANISOU 1261 OLEU A 203 5969 12777 4112 −2320 201 −371 A O ATOM 1262 CB LEU A 20339.199 12.266 23.416 1.00 75.30 A C ANISOU 1262 CB LEU A 203 7715 151845713 −2236 528 −808 A C ATOM 1263 CG LEU A 203 39.373 11.275 24.557 1.0086.53 A C ANISOU 1263 CG LEU A 203 9338 16113 7427 −2209 516 −1081 A CATOM 1264 CD1 LEU A 203 38.193 10.300 24.596 1.00 91.62 A C ANISOU 1264CD1 LEU A 203 10088 16741 7983 −2318 484 −1394 A C ATOM 1265 CD2 LEU A203 40.713 10.557 24.377 1.00 91.41 A C ANISOU 1265 CD2 LEU A 203 995816675 8101 −2124 639 −1302 A C ATOM 1266 N VAL A 204 38.641 14.75724.985 1.00 60.85 A N ANISOU 1266 N VAL A 204 5891 12955 4274 −2185 332−10 A N ATOM 1267 CA VAL A 204 38.563 15.562 26.200 1.00 58.10 A CANISOU 1267 CA VAL A 204 5608 12223 4247 −2147 237 247 A C ATOM 1268 CVAL A 204 37.214 16.262 26.276 1.00 57.43 A C ANISOU 1268 C VAL A 2045515 12228 4076 −2210 112 460 A C ATOM 1269 O VAL A 204 36.503 16.19427.293 1.00 63.08 A O ANISOU 1269 O VAL A 204 6351 12632 4984 −2234 1446 A O ATOM 1270 CB VAL A 204 39.730 16.570 26.248 1.00 57.99 A CANISOU 1270 CB VAL A 204 5486 12166 4383 −2067 311 534 A C ATOM 1271 CG1VAL A 204 39.459 17.645 27.289 1.00 55.56 A C ANISOU 1271 CG1 VAL A 2045200 11560 4350 −2044 212 853 A C ATOM 1272 CG2 VAL A 204 41.051 15.85026.507 1.00 58.13 A C ANISOU 1272 CG2 VAL A 204 5527 11999 4561 −1987405 295 A C ATOM 1273 N SER A 205 36.834 16.931 25.184 1.00 59.39 A NANISOU 1273 N SER A 205 5626 12916 4022 −2228 131 655 A N ATOM 1274 CASER A 205 35.606 17.716 25.205 1.00 58.96 A C ANISOU 1274 CA SER A 2055549 12978 3874 −2261 10 887 A C ATOM 1275 C SER A 205 34.384 16.83625.420 1.00 66.98 A C ANISOU 1275 C SER A 205 6650 13986 4814 −2350 −88586 A C ATOM 1276 O SER A 205 33.416 17.266 26.057 1.00 72.08 A O ANISOU1276 O SER A 205 7339 14501 5548 −2382 −214 702 A O ATOM 1277 CB SER A205 35.457 18.538 23.918 1.00 61.54 A C ANISOU 1277 CB SER A 205 572213807 3852 −2233 59 1142 A C ATOM 1278 OG SER A 205 35.619 17.748 22.7521.00 64.29 A O ANISOU 1278 OG SER A 205 6000 14513 3913 −2238 142 878 AO ATOM 1279 N SER A 206 34.407 15.602 24.920 1.00 70.00 A N ANISOU 1279N SER A 206 7054 14496 5049 −2399 −25 186 A N ATOM 1280 CA SER A 20633.249 14.742 25.127 1.00 66.46 A C ANISOU 1280 CA SER A 206 6681 140254547 −2501 −92 −129 A C ATOM 1281 C SER A 206 33.204 14.216 26.560 1.0057.36 A C ANISOU 1281 C SER A 206 5726 12306 3762 −2519 −129 −258 A CATOM 1282 O SER A 206 32.124 14.138 27.153 1.00 56.23 A O ANISOU 1282 OSER A 206 5654 12026 3684 −2592 −223 −313 A O ATOM 1283 CB SER A 20633.267 13.596 24.120 1.00 62.73 A C ANISOU 1283 CB SER A 206 6162 138673805 −2555 3 −529 A C ATOM 1284 OG SER A 206 34.365 12.747 24.396 1.0071.83 A O ANISOU 1284 OG SER A 206 7405 14763 5124 −2522 116 −746 A OATOM 1285 N LYS A 207 34.361 13.855 27.133 1.00 62.27 A N ANISOU 1285 NLYS A 207 6437 12602 4619 −2444 −54 −311 A N ATOM 1286 CA LYS A 20734.414 13.498 28.554 1.00 61.05 A C ANISOU 1286 CA LYS A 207 6478 119054815 −2420 −93 −376 A C ATOM 1287 C LYS A 207 33.789 14.580 29.418 1.0054.39 A C ANISOU 1287 C LYS A 207 5646 10871 4147 −2415 −228 −47 A CATOM 1288 O LYS A 207 32.980 14.295 30.304 1.00 57.02 A O ANISOU 1288 OLYS A 207 6113 10925 4626 −2466 −298 −132 A O ATOM 1289 CB LYS A 20735.861 13.255 29.006 1.00 56.97 A C ANISOU 1289 CB LYS A 207 6016 111204510 −2295 −12 −401 A C ATOM 1290 CG LYS A 207 36.315 11.831 28.819 1.0066.33 A C ANISOU 1290 CG LYS A 207 7305 12235 5664 −2291 102 −816 A CATOM 1291 CD LYS A 207 37.805 11.688 28.909 1.00 74.49 A C ANISOU 1291CD LYS A 207 8331 13150 6820 −2156 186 −835 A C ATOM 1292 CE LYS A 20738.233 10.435 28.141 1.00 83.59 A C ANISOU 1292 CE LYS A 207 9504 144487810 −2167 316 −1220 A C ATOM 1293 NZ LYS A 207 39.693 10.148 28.2521.00 87.90 A N1+ ANISOU 1293 NZ LYS A 207 10052 14869 8477 −2026 399−1293 A N1+ ATOM 1294 N PHE A 208 34.152 15.830 29.172 1.00 51.45 A NANISOU 1294 N PHE A 208 5138 10641 3768 −2358 −253 329 A N ATOM 1295 CAPHE A 208 33.703 16.910 30.035 1.00 52.14 A C ANISOU 1295 CA PHE A 2085234 10520 4055 −2341 −371 653 A C ATOM 1296 C PHE A 208 32.404 17.56229.583 1.00 56.75 A C ANISOU 1296 C PHE A 208 5734 11392 4435 −2412 −471808 A C ATOM 1297 O PHE A 208 31.812 18.318 30.356 1.00 51.09 A O ANISOU1297 O PHE A 208 5041 10489 3880 −2415 −582 1026 A O ATOM 1298 CB PHE A208 34.798 17.966 30.152 1.00 56.47 A C ANISOU 1298 CB PHE A 208 568911009 4758 −2241 −334 979 A C ATOM 1299 CG PHE A 208 35.953 17.52431.004 1.00 59.44 A C ANISOU 1299 CG PHE A 208 6154 11010 5421 −2150−285 861 A C ATOM 1300 CD1 PHE A 208 36.950 16.711 30.484 1.00 66.07 A CANISOU 1300 CD1 PHE A 208 6986 11926 6191 −2107 −161 625 A C ATOM 1301CD2 PHE A 208 36.010 17.868 32.347 1.00 55.08 A C ANISOU 1301 CD2 PHE A208 5694 10034 5199 −2096 −368 962 A C ATOM 1302 CE1 PHE A 208 38.00816.280 31.284 1.00 57.89 A C ANISOU 1302 CE1 PHE A 208 6031 10556 5409−1999 −125 499 A C ATOM 1303 CE2 PHE A 208 37.058 17.448 33.146 1.0051.66 A C ANISOU 1303 CE2 PHE A 208 5340 9278 5012 −1985 −333 839 A CATOM 1304 CZ PHE A 208 38.061 16.649 32.614 1.00 45.61 A C ANISOU 1304CZ PHE A 208 4564 8596 4171 −1932 −213 606 A C ATOM 1305 N GLY A 20931.901 17.174 28.418 1.00 66.58 A N ANISOU 1305 N GLY A 209 6878 130935326 −2460 −441 690 A N ATOM 1306 CA GLY A 209 30.677 17.751 27.897 1.0059.09 A C ANISOU 1306 CA GLY A 209 5836 12463 4151 −2502 −543 819 A CATOM 1307 C GLY A 209 30.813 19.235 27.648 1.00 55.88 A C ANISOU 1307 CGLY A 209 5316 12148 3769 −2404 −562 1280 A C ATOM 1308 O GLY A 20929.890 20.013 27.871 1.00 52.05 A O ANISOU 1308 O GLY A 209 4817 115163445 −2374 −645 1461 A O ATOM 1309 N ALA A 210 31.993 19.620 27.184 1.0064.40 A N ANISOU 1309 N ALA A 210 6311 13395 4764 −2333 −450 1442 A NATOM 1310 CA ALA A 210 32.298 21.003 26.868 1.00 58.70 A C ANISOU 1310CA ALA A 210 5501 12602 4201 −2223 −435 1846 A C ATOM 1311 C ALA A 21032.527 21.082 25.371 1.00 57.43 A C ANISOU 1311 C ALA A 210 5196 127793847 −2122 −379 1958 A C ATOM 1312 O ALA A 210 33.151 20.198 24.790 1.0057.64 A O ANISOU 1312 O ALA A 210 5164 12735 4001 −2020 −390 2258 A OATOM 1313 CB ALA A 210 33.532 21.465 27.623 1.00 58.08 A C ANISOU 1313CB ALA A 210 5447 12330 4290 −2206 −333 1985 A C ATOM 1314 N SER A 21132.001 22.122 24.739 1.00 59.87 A N ANISOU 1314 N SER A 211 5453 134323861 −2139 −322 1706 A N ATOM 1315 CA SER A 211 32.158 22.270 23.3001.00 76.19 A C ANISOU 1315 CA SER A 211 7393 15848 5708 −2037 −255 1787A C ATOM 1316 C SER A 211 33.620 22.433 22.910 1.00 80.59 A C ANISOU1316 C SER A 211 7920 16415 6287 −1973 −123 1979 A C ATOM 1317 O SER A211 34.369 23.155 23.565 1.00 79.45 A O ANISOU 1317 O SER A 211 781415975 6397 −1957 −101 2200 A O ATOM 1318 CB SER A 211 31.374 23.48722.809 1.00 87.70 A C ANISOU 1318 CB SER A 211 8782 17365 7174 −1931−325 2014 A C ATOM 1319 OG SER A 211 31.911 24.684 23.350 1.00 86.65 A OANISOU 1319 OG SER A 211 8548 17571 6805 −1820 −253 2092 A O ATOM 1320 NGLN A 212 34.018 21.774 21.825 1.00 94.03 A N ANISOU 1320 N GLN A 2129542 18451 7732 −1932 −28 1886 A N ATOM 1321 CA GLN A 212 35.389 21.87021.334 1.00 73.86 A C ANISOU 1321 CA GLN A 212 6963 15934 5166 −1899 1261967 A C ATOM 1322 C GLN A 212 35.714 23.297 20.906 1.00 68.66 A CANISOU 1322 C GLN A 212 6280 15163 4645 −1784 174 2363 A C ATOM 1323 OGLN A 212 36.815 23.783 21.131 1.00 68.36 A O ANISOU 1323 O GLN A 2126250 14967 4756 −1776 291 2483 A O ATOM 1324 CB GLN A 212 35.668 20.86220.216 1.00 71.09 A C ANISOU 1324 CB GLN A 212 6538 15973 4501 −1891 2041720 A C ATOM 1325 CG GLN A 212 34.441 20.368 19.476 1.00 70.66 A CANISOU 1325 CG GLN A 212 6512 15998 4338 −2000 146 1295 A C ATOM 1326 CDGLN A 212 34.676 19.025 18.811 1.00 78.81 A C ANISOU 1326 CD GLN A 2127458 17398 5090 −1994 207 1005 A C ATOM 1327 OE1 GLN A 212 35.773 18.47418.874 1.00 87.67 A O ANISOU 1327 OE1 GLN A 212 8576 18575 6161 −2020324 837 A O ATOM 1328 NE2 GLN A 212 33.642 18.489 18.174 1.00 75.62 A NANISOU 1328 NE2 GLN A 212 6974 17243 4515 −1958 131 923 A N ATOM 1329 NVAL A 213 34.753 23.964 20.283 1.00 74.66 A N ANISOU 1329 N VAL A 2137016 15968 5385 −1696 96 2550 A N ATOM 1330 CA VAL A 213 34.957 25.33719.862 1.00 70.77 A C ANISOU 1330 CA VAL A 213 6512 15397 4982 −1574 1672895 A C ATOM 1331 C VAL A 213 35.206 26.206 21.088 1.00 68.00 A CANISOU 1331 C VAL A 213 6220 14608 5008 −1573 130 3102 A C ATOM 1332 OVAL A 213 36.042 27.103 21.053 1.00 68.58 A O ANISOU 1332 O VAL A 2136302 14520 5234 −1504 230 3338 A O ATOM 1333 CB VAL A 213 33.751 25.87719.078 1.00 73.33 A C ANISOU 1333 CB VAL A 213 6786 15984 5092 −1460 1192982 A C ATOM 1334 CG1 VAL A 213 33.995 27.318 18.658 1.00 74.75 A CANISOU 1334 CG1 VAL A 213 6982 16058 5362 −1324 191 3337 A C ATOM 1335CG2 VAL A 213 33.479 25.001 17.868 1.00 76.22 A C ANISOU 1335 CG2 VAL A213 7085 16763 5113 −1452 175 2773 A C ATOM 1336 N GLU A 214 34.47325.951 22.168 1.00 68.86 A N ANISOU 1336 N GLU A 214 6373 14511 5281−1647 −8 3006 A N ATOM 1337 CA GLU A 214 34.675 26.729 23.386 1.00 65.76A C ANISOU 1337 CA GLU A 214 6029 13698 5259 −1638 −57 3175 A C ATOM1338 C GLU A 214 36.075 26.501 23.938 1.00 64.27 A C ANISOU 1338 C GLU A214 5869 13285 5266 −1685 39 3157 A C ATOM 1339 O GLU A 214 36.77127.446 24.334 1.00 69.95 A O ANISOU 1339 O GLU A 214 6595 13743 6241−1628 99 3344 A O ATOM 1340 CB GLU A 214 33.613 26.382 24.424 1.00 71.17A C ANISOU 1340 CB GLU A 214 6753 14234 6053 −1700 −226 3070 A C ATOM1341 CG GLU A 214 32.237 26.915 24.072 1.00 85.00 A C ANISOU 1341 CG GLUA 214 8465 16141 7690 −1633 −303 3123 A C ATOM 1342 CD GLU A 214 32.22428.424 23.942 1.00 98.77 A C ANISOU 1342 CD GLU A 214 10194 17748 9586−1500 −273 3417 A C ATOM 1343 OE1 GLU A 214 33.167 29.069 24.445 1.0098.79 A O ANISOU 1343 OE1 GLU A 214 10225 17456 9854 −1475 −224 3561 A OATOM 1344 OE2 GLU A 214 31.274 28.969 23.342 1.00 104.66 A O1− ANISOU1344 OE2 GLU A 214 10904 18679 10181 −1415 −293 3485 A O1− ATOM 1345 NMET A 215 36.509 25.245 23.910 1.00 61.93 A N ANISOU 1345 N MET A 2155586 13093 4852 −1784 72 2906 A N ATOM 1346 CA MET A 215 37.833 24.90524.389 1.00 60.37 A C ANISOU 1346 CA MET A 215 5382 12780 4775 −1815 2132856 A C ATOM 1347 C MET A 215 38.867 25.617 23.547 1.00 62.68 A CANISOU 1347 C MET A 215 5618 13111 5084 −1731 373 3048 A C ATOM 1348 OMET A 215 39.794 26.221 24.077 1.00 65.61 A O ANISOU 1348 O MET A 2155993 13190 5747 −1697 433 3195 A O ATOM 1349 CB MET A 215 38.081 23.41224.229 1.00 62.60 A C ANISOU 1349 CB MET A 215 5659 13308 4817 −1905 2772537 A C ATOM 1350 CG MET A 215 37.535 22.534 25.330 1.00 66.60 A CANISOU 1350 CG MET A 215 6249 13701 5354 −2006 175 2318 A C ATOM 1351 SDMET A 215 38.234 20.888 25.148 1.00 72.76 A S ANISOU 1351 SD MET A 2157044 14664 5937 −2078 282 1882 A S ATOM 1352 CE MET A 215 39.973 21.24825.348 1.00 67.92 A C ANISOU 1352 CE MET A 215 6385 13768 5652 −2025 4251881 A C ATOM 1353 N LEU A 216 38.677 25.593 22.231 1.00 65.71 A NANISOU 1353 N LEU A 216 5957 13852 5158 −1695 447 3038 A N ATOM 1354 CALEU A 216 39.630 26.233 21.341 1.00 68.27 A C ANISOU 1354 CA LEU A 2166241 14243 5455 −1634 632 3185 A C ATOM 1355 C LEU A 216 39.696 27.72921.597 1.00 68.48 A C ANISOU 1355 C LEU A 216 6295 14024 5701 −1543 6513491 A C ATOM 1356 O LEU A 216 40.772 28.314 21.598 1.00 72.05 A OANISOU 1356 O LEU A 216 6740 14320 6317 −1524 811 3605 A O ATOM 1357 CBLEU A 216 39.279 25.961 19.879 1.00 80.18 A C ANISOU 1357 CB LEU A 2167706 16187 6573 −1601 684 3108 A C ATOM 1358 CG LEU A 216 39.370 24.50919.407 1.00 85.93 A C ANISOU 1358 CG LEU A 216 8411 17058 7182 −1703 7072767 A C ATOM 1359 CD1 LEU A 216 38.850 24.367 17.985 1.00 78.33 A CANISOU 1359 CD1 LEU A 216 7395 16514 5851 −1690 758 2590 A C ATOM 1360CD2 LEU A 216 40.790 23.985 19.522 1.00 99.52 A C ANISOU 1360 CD2 LEU A216 10111 18564 9138 −1743 859 2736 A C ATOM 1361 N ALA A 217 38.54628.350 21.821 1.00 67.78 A N ANISOU 1361 N ALA A 217 6235 13882 5637−1492 502 3599 A N ATOM 1362 CA ALA A 217 38.524 29.780 22.062 1.0067.65 A C ANISOU 1362 CA ALA A 217 6251 13588 5864 −1408 513 3849 A CATOM 1363 C ALA A 217 39.286 30.106 23.329 1.00 65.09 A C ANISOU 1363 CALA A 217 5948 12855 5929 −1444 534 3850 A C ATOM 1364 O ALA A 21740.053 31.061 23.376 1.00 65.90 A O ANISOU 1364 O ALA A 217 6060 127616219 −1404 667 3995 A O ATOM 1365 CB ALA A 217 37.092 30.272 22.170 1.0066.83 A C ANISOU 1365 CB ALA A 217 6162 13476 5755 −1357 335 3908 A CATOM 1366 N GLU A 218 39.055 29.308 24.363 1.00 62.20 A N ANISOU 1366 NGLU A 218 5594 12356 5684 −1520 416 3673 A N ATOM 1367 CA GLU A 21839.727 29.504 25.642 1.00 63.47 A C ANISOU 1367 CA GLU A 218 5773 121216221 −1532 412 3659 A C ATOM 1368 C GLU A 218 41.237 29.262 25.640 1.0075.47 A C ANISOU 1368 C GLU A 218 7246 13603 7827 −1574 603 3574 A CATOM 1369 O GLU A 218 41.990 29.982 26.293 1.00 75.19 A O ANISOU 1369 OGLU A 218 7203 13262 8104 −1557 664 3601 A O ATOM 1370 CB GLU A 21839.069 28.655 26.731 1.00 56.56 A C ANISOU 1370 CB GLU A 218 4941 111025447 −1584 220 3509 A C ATOM 1371 CG GLU A 218 39.656 28.867 28.116 1.0061.42 A C ANISOU 1371 CG GLU A 218 5582 11321 6434 −1580 189 3460 A CATOM 1372 CD GLU A 218 39.389 30.254 28.654 1.00 72.09 A C ANISOU 1372CD GLU A 218 6947 12385 8057 −1485 176 3620 A C ATOM 1373 OE1 GLU A 21840.314 30.863 29.225 1.00 80.76 A O ANISOU 1373 OE1 GLU A 218 8018 133259341 −1464 320 3652 A O ATOM 1374 OE2 GLU A 218 38.252 30.739 28.5031.00 69.14 A O1− ANISOU 1374 OE2 GLU A 218 6602 11949 7718 −1440 31 3686A O1− ATOM 1375 N ASN A 219 41.674 28.230 24.932 1.00 78.62 A N ANISOU1375 N ASN A 219 7598 14314 7959 −1625 705 3445 A N ATOM 1376 CA ASN A219 43.082 27.844 24.923 1.00 73.26 A C ANISOU 1376 CA ASN A 219 684913623 7362 −1675 871 3295 A C ATOM 1377 C ASN A 219 43.973 28.265 23.7561.00 82.74 A C ANISOU 1377 C ASN A 219 7998 14985 8455 −1657 1097 3372 AC ATOM 1378 O ASN A 219 45.178 28.059 23.830 1.00 95.69 A O ANISOU 1378O ASN A 219 9563 16580 10216 −1694 1252 3253 A O ATOM 1379 CB ASN A 21943.181 26.326 25.058 1.00 64.39 A C ANISOU 1379 CB ASN A 219 5701 127206043 −1753 841 3022 A C ATOM 1380 CG ASN A 219 42.799 25.840 26.432 1.0058.78 A C ANISOU 1380 CG ASN A 219 5035 11780 5517 −1793 684 2897 A CATOM 1381 OD1 ASN A 219 43.374 26.260 27.428 1.00 60.24 A O ANISOU 1381OD1 ASN A 219 5185 11706 5997 −1799 711 2815 A O ATOM 1382 ND2 ASN A 21941.831 24.943 26.491 1.00 57.87 A N ANISOU 1382 ND2 ASN A 219 4992 117605234 −1819 523 2867 A N ATOM 1383 N ASN A 220 43.396 28.809 22.687 1.0080.02 A N ANISOU 1383 N ASN A 220 7685 14828 7890 −1598 1124 3552 A NATOM 1384 CA ASN A 220 44.159 29.235 21.499 1.00 82.61 A C ANISOU 1384CA ASN A 220 7983 15376 8029 −1579 1334 3621 A C ATOM 1385 C ASN A 22045.146 28.165 20.998 1.00 77.17 A C ANISOU 1385 C ASN A 220 7207 148687246 −1648 1475 3383 A C ATOM 1386 O ASN A 220 46.310 28.433 20.732 1.0073.61 A O ANISOU 1386 O ASN A 220 6699 14304 6965 −1674 1665 3359 A OATOM 1387 CB ASN A 220 44.693 30.693 21.538 1.00 88.14 A C ANISOU 1387CB ASN A 220 8712 15823 8953 −1535 1487 3852 A C ATOM 1388 CG ASN A 22045.834 30.908 22.513 1.00 90.15 A C ANISOU 1388 CG ASN A 220 8914 157579582 −1590 1619 3771 A C ATOM 1389 OD1 ASN A 220 46.845 30.211 22.4791.00 92.93 A O ANISOU 1389 OD1 ASN A 220 9217 15997 10095 −1637 15463576 A O ATOM 1390 ND2 ASN A 220 45.683 31.902 23.378 1.00 84.96 A NANISOU 1390 ND2 ASN A 220 8266 14952 9063 −1582 1820 3909 A N ATOM 1391N HIS A 221 44.638 26.942 20.889 1.00 75.95 A N ANISOU 1391 N HIS A 2217037 15017 6802 −1676 1390 3185 A N ATOM 1392 CA HIS A 221 45.401 25.77620.453 1.00 72.96 A C ANISOU 1392 CA HIS A 221 6582 14812 6327 −17401466 2884 A C ATOM 1393 C HIS A 221 46.700 25.489 21.176 1.00 71.64 A CANISOU 1393 C HIS A 221 6339 14394 6488 −1782 1571 2760 A C ATOM 1394 OHIS A 221 47.742 25.322 20.555 1.00 75.41 A O ANISOU 1394 O HIS A 2216752 14848 7051 −1783 1767 2783 A O ATOM 1395 CB HIS A 221 45.612 25.68218.950 1.00 80.28 A C ANISOU 1395 CB HIS A 221 7463 16072 6966 −17271628 2834 A C ATOM 1396 CG HIS A 221 46.214 24.377 18.536 1.00 79.83 A CANISOU 1396 CG HIS A 221 7346 16262 6722 −1778 1641 2483 A C ATOM 1397ND1 HIS A 221 45.895 23.190 19.160 1.00 75.02 A N ANISOU 1397 ND1 HIS A221 6767 15744 5991 −1813 1476 2269 A N ATOM 1398 CD2 HIS A 221 47.12424.068 17.584 1.00 81.45 A C ANISOU 1398 CD2 HIS A 221 7467 16625 6855−1801 1805 2285 A C ATOM 1399 CE1 HIS A 221 46.576 22.205 18.605 1.0075.67 A C ANISOU 1399 CE1 HIS A 221 6795 16027 5930 −1850 1541 1949 A CATOM 1400 NE2 HIS A 221 47.328 22.711 17.645 1.00 78.84 A N ANISOU 1400NE2 HIS A 221 7120 16475 6359 −1838 1732 1952 A N ATOM 1401 N ASN A 22246.660 25.377 22.493 1.00 68.73 A N ANISOU 1401 N ASN A 222 5966 138536295 −1816 1452 2604 A N ATOM 1402 CA ASN A 222 47.888 25.104 23.2171.00 80.29 A C ANISOU 1402 CA ASN A 222 7324 15174 8009 −1842 1543 2393A C ATOM 1403 C ASN A 222 48.243 23.616 23.233 1.00 92.14 A C ANISOU1403 C ASN A 222 8757 16963 9290 −1876 1597 2052 A C ATOM 1404 O ASN A222 49.369 23.240 23.556 1.00 103.64 A O ANISOU 1404 O ASN A 222 1009618380 10903 −1879 1710 1843 A O ATOM 1405 CB ASN A 222 47.808 25.65424.643 1.00 98.25 A C ANISOU 1405 CB ASN A 222 9618 17095 10615 −18401396 2385 A C ATOM 1406 CG ASN A 222 49.181 25.937 25.238 1.00 113.99 AC ANISOU 1406 CG ASN A 222 11580 18733 12999 −1805 1457 2539 A C ATOM1407 OD1 ASN A 222 50.210 25.565 24.668 1.00 116.43 A O ANISOU 1407 OD1ASN A 222 11767 18922 13551 −1804 1563 2384 A O ATOM 1408 ND2 ASN A 22249.202 26.601 26.386 1.00 116.50 A N ANISOU 1408 ND2 ASN A 222 1199818890 13375 −1770 1391 2809 A N ATOM 1409 N PHE A 223 47.294 22.78222.834 1.00 87.72 A N ANISOU 1409 N PHE A 223 8260 16704 8366 −1890 15311971 A N ATOM 1410 CA PHE A 223 47.448 21.321 22.858 1.00 82.07 A CANISOU 1410 CA PHE A 223 7553 16124 7506 −1915 1483 1602 A C ATOM 1411 CPHE A 223 48.659 20.641 22.211 1.00 77.37 A C ANISOU 1411 C PHE A 2236875 15736 6785 −1904 1630 1312 A C ATOM 1412 O PHE A 223 49.150 19.65322.747 1.00 70.48 A O ANISOU 1412 O PHE A 223 6062 14768 5949 −1865 1553939 A O ATOM 1413 CB PHE A 223 46.174 20.672 22.311 1.00 81.91 A CANISOU 1413 CB PHE A 223 7637 16310 7175 −1940 1343 1589 A C ATOM 1414CG PHE A 223 44.924 21.132 22.997 1.00 78.81 A C ANISOU 1414 CG PHE A223 7330 15706 6908 −1949 1178 1819 A C ATOM 1415 CD1 PHE A 223 44.35220.377 24.002 1.00 73.06 A C ANISOU 1415 CD1 PHE A 223 6697 14567 6494−1924 1023 1682 A C ATOM 1416 CD2 PHE A 223 44.325 22.326 22.640 1.0083.58 A C ANISOU 1416 CD2 PHE A 223 7985 16361 7411 −1906 1103 2108 A CATOM 1417 CE1 PHE A 223 43.202 20.802 24.635 1.00 81.86 A C ANISOU 1417CE1 PHE A 223 7887 15490 7726 −1936 873 1888 A C ATOM 1418 CE2 PHE A 22343.176 22.759 23.268 1.00 71.01 A C ANISOU 1418 CE2 PHE A 223 6472 145525957 −1899 938 2293 A C ATOM 1419 CZ PHE A 223 42.613 21.996 24.267 1.0074.79 A C ANISOU 1419 CZ PHE A 223 6988 14762 6668 −1944 850 2210 A CATOM 1420 N THR A 224 49.140 21.116 21.073 1.00 85.28 A N ANISOU 1420 NTHR A 224 7792 16921 7691 −1902 1806 1438 A N ATOM 1421 CA THR A 22450.284 20.441 20.460 1.00 91.47 A C ANISOU 1421 CA THR A 224 8474 178828399 −1900 1955 1139 A C ATOM 1422 C THR A 224 51.539 20.448 21.337 1.0093.58 A C ANISOU 1422 C THR A 224 8645 17868 9045 −1871 2016 989 A CATOM 1423 O THR A 224 52.255 19.452 21.403 1.00 95.06 A O ANISOU 1423 OTHR A 224 8798 18057 9262 −1825 2043 635 A O ATOM 1424 CB THR A 22450.640 21.024 19.079 1.00 84.32 A C ANISOU 1424 CB THR A 224 7541 172007298 −1886 2100 1264 A C ATOM 1425 OG1 THR A 224 50.570 22.454 19.1271.00 80.96 A O ANISOU 1425 OG1 THR A 224 7151 16617 6993 −1870 2137 1657A O ATOM 1426 CG2 THR A 224 49.686 20.506 18.021 1.00 80.74 A C ANISOU1426 CG2 THR A 224 7144 17094 6438 −1882 2037 1163 A C ATOM 1427 N ALA A225 51.810 21.568 21.997 1.00 90.56 A N ANISOU 1427 N ALA A 225 823117206 8972 −1874 2013 1226 A N ATOM 1428 CA ALA A 225 52.997 21.68422.844 1.00 92.73 A C ANISOU 1428 CA ALA A 225 8390 17230 9611 −18352084 1092 A C ATOM 1429 C ALA A 225 52.780 21.585 24.358 1.00 83.47 A CANISOU 1429 C ALA A 225 7293 15633 8788 −1759 1881 1006 A C ATOM 1430 OALA A 225 53.715 21.804 25.124 1.00 90.51 A O ANISOU 1430 O ALA A 2258082 16302 10006 −1713 1917 913 A O ATOM 1431 CB ALA A 225 53.750 22.96422.510 1.00 96.06 A C ANISOU 1431 CB ALA A 225 8739 17582 10176 −18652262 1372 A C ATOM 1432 N SER A 226 51.568 21.269 24.798 1.00 72.08 A NANISOU 1432 N SER A 226 6022 14081 7286 −1742 1673 1015 A N ATOM 1433 CASER A 226 51.296 21.208 26.238 1.00 64.73 A C ANISOU 1433 CA SER A 2265168 12748 6676 −1677 1493 1008 A C ATOM 1434 C SER A 226 51.233 19.84626.918 1.00 62.92 A C ANISOU 1434 C SER A 226 5097 12346 6463 −1579 1318652 A C ATOM 1435 O SER A 226 50.841 19.770 28.076 1.00 60.54 A O ANISOU1435 O SER A 226 4930 11778 6295 −1543 1141 672 A O ATOM 1436 CB SER A226 50.026 21.985 26.585 1.00 63.08 A C ANISOU 1436 CB SER A 226 502912475 6463 −1740 1404 1378 A C ATOM 1437 OG SER A 226 48.901 21.41225.951 1.00 62.59 A O ANISOU 1437 OG SER A 226 5122 12523 6137 −17601268 1341 A O ATOM 1438 N THR A 227 51.600 18.767 26.244 1.00 64.35 A NANISOU 1438 N THR A 227 5271 12659 6519 −1531 1374 325 A N ATOM 1439 CATHR A 227 51.485 17.485 26.924 1.00 63.11 A C ANISOU 1439 CA THR A 2275277 12299 6403 −1422 1231 −14 A C ATOM 1440 C THR A 227 52.078 17.57728.331 1.00 61.15 A C ANISOU 1440 C THR A 227 5040 11662 6533 −1284 1136−110 A C ATOM 1441 O THR A 227 53.120 18.202 28.533 1.00 61.68 A OANISOU 1441 O THR A 227 4937 11695 6804 −1247 1226 −102 A O ATOM 1442 CBTHR A 227 52.178 16.413 26.097 1.00 65.51 A C ANISOU 1442 CB THR A 2275557 12824 6511 −1391 1329 −344 A C ATOM 1443 OG1 THR A 227 51.63116.424 24.760 1.00 67.26 A O ANISOU 1443 OG1 THR A 227 5748 13439 6370−1517 1421 −242 A O ATOM 1444 CG2 THR A 227 52.012 15.042 26.760 1.0064.24 A C ANISOU 1444 CG2 THR A 227 5588 12439 6380 −1274 1202 −691 A CATOM 1445 N ASN A 228 51.384 16.990 29.316 1.00 60.48 A N ANISOU 1445 NASN A 228 5151 11289 6541 −1207 959 −199 A N ATOM 1446 CA ASN A 22851.729 17.108 30.743 1.00 56.94 A C ANISOU 1446 CA ASN A 228 4742 104636430 −1061 841 −257 A C ATOM 1447 C ASN A 228 51.839 18.567 31.179 1.0056.10 A C ANISOU 1447 C ASN A 228 4483 10276 6555 −1110 855 53 A C ATOM1448 O ASN A 228 52.673 18.921 32.013 1.00 55.52 A O ANISOU 1448 O ASN A228 4315 10015 6765 −1001 840 −16 A O ATOM 1449 CB ASN A 228 53.01216.329 31.087 1.00 57.75 A C ANISOU 1449 CB ASN A 228 4808 10483 6650−880 867 −615 A C ATOM 1450 CG ASN A 228 52.849 14.815 30.882 1.00 71.18A C ANISOU 1450 CG ASN A 228 6699 12180 8166 −804 838 −934 A C ATOM 1451OD1 ASN A 228 51.733 14.296 30.974 1.00 72.60 A O ANISOU 1451 OD1 ASN A228 7076 12284 8226 −852 755 −917 A O ATOM 1452 ND2 ASN A 228 53.96914.104 30.608 1.00 99.89 A N ANISOU 1452 ND2 ASN A 228 10272 15894 11787−689 917 −1238 A N ATOM 1453 N ARC A 229 51.023 19.424 30.571 1.00 56.26A N ANISOU 1453 N ARC A 229 4472 10455 6449 −1269 894 384 A N ATOM 1454CA ARC A 229 50.807 20.794 30.998 1.00 55.28 A C ANISOU 1454 CA ARC A229 4252 10227 6525 −1330 895 716 A C ATOM 1455 C ARC A 229 49.31021.022 31.159 1.00 53.69 A C ANISOU 1455 C ARC A 229 4202 9976 6224−1409 763 952 A C ATOM 1456 O ARC A 229 48.476 20.320 30.583 1.00 54.03A O ANISOU 1456 O ARC A 229 4371 10168 5991 −1458 723 903 A O ATOM 1457CB ARC A 229 51.378 21.812 30.015 1.00 57.50 A C ANISOU 1457 CB ARC A229 4327 10763 6757 −1442 1111 931 A C ATOM 1458 CG ARC A 229 52.86821.938 30.115 1.00 62.06 A C ANISOU 1458 CG ARC A 229 4715 11328 7535−1379 1243 741 A C ATOM 1459 CD ARC A 229 53.415 23.161 29.386 1.0063.73 A C ANISOU 1459 CD ARC A 229 4721 11708 7784 −1500 1474 990 A CATOM 1460 NE ARC A 229 54.770 23.465 29.843 1.00 61.42 A N ANISOU 1460NE ARC A 229 4232 11318 7787 −1441 1575 809 A N ATOM 1461 CZ ARC A 22955.861 22.800 29.463 1.00 75.74 A C ANISOU 1461 CZ ARC A 229 5947 132619569 −1381 1672 496 A C ATOM 1462 NH1 ARC A 229 55.763 21.795 28.5991.00 75.41 A N1+ ANISOU 1462 NH1 ARC A 229 5992 13448 9212 −1379 1688341 A N1+ ATOM 1463 NH2 ARC A 229 57.051 23.146 29.946 1.00 78.66 A NANISOU 1463 NH2 ARC A 229 6120 13542 10225 −1325 1754 323 A N ATOM 1464N SER A 230 48.955 21.974 32.006 1.00 56.81 A N ANISOU 1464 N SER A 2304568 10167 6850 −1423 698 1194 A N ATOM 1465 CA SER A 230 47.556 22.21432.290 1.00 50.24 A C ANISOU 1465 CA SER A 230 3879 9234 5978 −1475 5481387 A C ATOM 1466 C SER A 230 46.873 23.208 31.375 1.00 51.39 A CANISOU 1466 C SER A 230 3952 9627 5945 −1612 635 1751 A C ATOM 1467 OSER A 230 47.416 24.257 31.060 1.00 52.25 A O ANISOU 1467 O SER A 2303905 9769 6178 −1653 765 1974 A O ATOM 1468 CB SER A 230 47.382 22.63133.743 1.00 47.73 A C ANISOU 1468 CB SER A 230 3587 8542 6005 −1396 4091430 A C ATOM 1469 OG SER A 230 47.887 23.922 33.939 1.00 54.39 A OANISOU 1469 OG SER A 230 4278 9370 7019 −1464 478 1739 A O ATOM 1470 NVAL A 231 45.649 22.857 30.995 1.00 51.59 A N ANISOU 1470 N VAL A 2314095 9830 5677 −1676 572 1795 A N ATOM 1471 CA VAL A 231 44.806 23.64730.113 1.00 52.52 A C ANISOU 1471 CA VAL A 231 4191 10190 5574 −1775 6032122 A C ATOM 1472 C VAL A 231 43.419 23.784 30.724 1.00 50.28 A CANISOU 1472 C VAL A 231 4034 9736 5335 −1795 411 2251 A C ATOM 1473 OVAL A 231 43.089 23.094 31.679 1.00 48.05 A O ANISOU 1473 O VAL A 2313850 9137 5268 −1739 272 2104 A O ATOM 1474 CB VAL A 231 44.691 23.01728.719 1.00 54.92 A C ANISOU 1474 CB VAL A 231 4505 10895 5469 −1819 6832023 A C ATOM 1475 CG1 VAL A 231 45.988 23.187 27.953 1.00 57.44 A CANISOU 1475 CG1 VAL A 231 4676 11428 5722 −1819 900 1989 A C ATOM 1476CG2 VAL A 231 44.319 21.552 28.829 1.00 54.30 A C ANISOU 1476 CG2 VAL A231 4574 10777 5281 −1791 572 1652 A C ATOM 1477 N LEU A 232 42.63524.719 30.207 1.00 51.03 A N ANISOU 1477 N LEU A 232 4127 10051 5209−1865 404 2521 A N ATOM 1478 CA LEU A 232 41.290 24.966 30.700 1.0049.17 A C ANISOU 1478 CA LEU A 232 4017 9634 5033 −1849 213 2649 A CATOM 1479 C LEU A 232 40.163 24.390 29.846 1.00 50.12 A C ANISOU 1479 CLEU A 232 4196 10067 4782 −1913 152 2640 A C ATOM 1480 O LEU A 23240.169 24.506 28.632 1.00 63.63 A O ANISOU 1480 O LEU A 232 5866 120816229 −1899 227 2706 A O ATOM 1481 CB LEU A 232 41.078 26.464 30.816 1.0049.13 A C ANISOU 1481 CB LEU A 232 4026 9422 5219 −1743 186 2886 A CATOM 1482 CG LEU A 232 42.043 27.197 31.732 1.00 47.52 A C ANISOU 1482CG LEU A 232 3810 8827 5417 −1664 180 2887 A C ATOM 1483 CD1 LEU A 23242.076 28.667 31.371 1.00 51.45 A C ANISOU 1483 CD1 LEU A 232 4299 92256023 −1582 228 3085 A C ATOM 1484 CD2 LEU A 232 41.611 27.012 33.1721.00 44.54 A C ANISOU 1484 CD2 LEU A 232 3541 8151 5232 −1635 −13 2826 AC ATOM 1485 N ILE A 233 39.205 23.754 30.513 1.00 48.36 A N ANISOU 1485N ILE A 233 4081 9738 4555 −1957 3 2525 A N ATOM 1486 CA ILE A 23338.008 23.176 29.913 1.00 49.07 A C ANISOU 1486 CA ILE A 233 4238 100784328 −2013 −77 2431 A C ATOM 1487 C ILE A 233 36.832 24.078 30.282 1.0051.00 A C ANISOU 1487 C ILE A 233 4519 10221 4638 −1994 −235 2650 A CATOM 1488 O ILE A 233 36.416 24.092 31.453 1.00 45.43 A O ANISOU 1488 OILE A 233 3896 9193 4172 −1998 −356 2637 A O ATOM 1489 CB ILE A 23337.753 21.740 30.396 1.00 48.14 A C ANISOU 1489 CB ILE A 233 4265 98104216 −2023 −153 2024 A C ATOM 1490 CG1 ILE A 233 38.602 20.702 29.6901.00 49.95 A C ANISOU 1490 CG1 ILE A 233 4485 10197 4298 −2006 −32 1723A C ATOM 1491 CG2 ILE A 233 36.364 21.331 30.050 1.00 48.44 A C ANISOU1491 CG2 ILE A 233 4363 10034 4007 −2096 −253 1945 A C ATOM 1492 CD1 ILEA 233 40.039 21.006 29.647 1.00 50.56 A C ANISOU 1492 CD1 ILE A 233 446410219 4526 −1940 100 1756 A C ATOM 1493 N PRO A 234 36.271 24.863 29.3411.00 50.40 A N ANISOU 1493 N PRO A 234 4388 10352 4409 −1931 −256 2796 AN ATOM 1494 CA PRO A 234 35.116 25.691 29.696 1.00 49.30 A C ANISOU 1494CA PRO A 234 4264 10082 4387 −1881 −412 2915 A C ATOM 1495 C PRO A 23433.828 24.891 29.617 1.00 51.11 A C ANISOU 1495 C PRO A 234 4534 104774408 −1975 −514 2755 A C ATOM 1496 O PRO A 234 33.408 24.463 28.530 1.0051.09 A O ANISOU 1496 O PRO A 234 4486 10838 4086 −1999 −477 2666 A OATOM 1497 CB PRO A 234 35.144 26.813 28.649 1.00 51.01 A C ANISOU 1497CB PRO A 234 4388 10460 4532 −1776 −347 3114 A C ATOM 1498 CG PRO A 23436.390 26.584 27.853 1.00 53.06 A C ANISOU 1498 CG PRO A 234 4603 108944664 −1772 −165 3110 A C ATOM 1499 CD PRO A 234 36.704 25.143 27.9701.00 52.71 A C ANISOU 1499 CD PRO A 234 4590 10969 4467 −1886 −126 2860A C ATOM 1500 N VAL A 235 33.207 24.699 30.783 1.00 50.00 A N ANISOU1500 N VAL A 235 4482 10053 4461 −2019 −639 2696 A N ATOM 1501 CA VAL A235 32.033 23.853 30.916 1.00 52.81 A C ANISOU 1501 CA VAL A 235 490510495 4664 −2128 −721 2506 A C ATOM 1502 C VAL A 235 30.898 24.63631.573 1.00 52.27 A C ANISOU 1502 C VAL A 235 4832 10207 4823 −2103 −8352581 A C ATOM 1503 O VAL A 235 31.109 25.398 32.533 1.00 62.35 A OANISOU 1503 O VAL A 235 6117 11143 6431 −2034 −897 2689 A O ATOM 1504 CBVAL A 235 32.366 22.559 31.692 1.00 49.50 A C ANISOU 1504 CB VAL A 2354641 9944 4224 −2236 −720 2285 A C ATOM 1505 CG1 VAL A 235 32.413 22.79433.193 1.00 41.14 A C ANISOU 1505 CG1 VAL A 235 3676 8401 3555 −2216−807 2332 A C ATOM 1506 CG2 VAL A 235 31.382 21.471 31.327 1.00 62.52 AC ANISOU 1506 CG2 VAL A 235 6351 11764 5639 −2320 −765 1954 A C ATOM1507 N THR A 236 29.695 24.471 31.008 1.00 53.61 A N ANISOU 1507 N THR A236 4981 10579 4808 −2141 −845 2491 A N ATOM 1508 CA THR A 236 28.50125.154 31.505 1.00 65.00 A C ANISOU 1508 CA THR A 236 6426 11851 6420−2127 −901 2538 A C ATOM 1509 C THR A 236 28.045 24.560 32.827 1.0075.05 A C ANISOU 1509 C THR A 236 7859 12779 7876 −2215 −955 2410 A CATOM 1510 O THR A 236 27.655 25.286 33.753 1.00 79.19 A O ANISOU 1510 OTHR A 236 8419 12979 8690 −2173 −992 2482 A O ATOM 1511 CB THR A 23627.379 25.032 30.477 1.00 60.90 A C ANISOU 1511 CB THR A 236 5859 116625619 −2122 −881 2452 A C ATOM 1512 OG1 THR A 236 27.306 23.658 30.0691.00 67.49 A O ANISOU 1512 OG1 THR A 236 6751 12709 6182 −2214 −864 2174A O ATOM 1513 CG2 THR A 236 27.648 25.925 29.273 1.00 55.56 A C ANISOU1513 CG2 THR A 236 5041 11260 4809 −1999 −830 2635 A C ATOM 1514 N SER A237 28.046 23.236 32.914 1.00 70.58 A N ANISOU 1514 N SER A 237 742312258 7135 −2305 −955 2172 A N ATOM 1515 CA SER A 237 27.749 22.54334.157 1.00 68.98 A C ANISOU 1515 CA SER A 237 7428 11694 7086 −2351−1004 2024 A C ATOM 1516 C SER A 237 28.769 21.429 34.394 1.00 60.64 A CANISOU 1516 C SER A 237 6451 10597 5992 −2394 −1025 1828 A C ATOM 1517 OSER A 237 29.389 20.915 33.462 1.00 56.52 A O ANISOU 1517 O SER A 2375844 10400 5231 −2429 −989 1726 A O ATOM 1518 CB SER A 237 26.331 21.96834.133 1.00 65.84 A C ANISOU 1518 CB SER A 237 7116 11322 6579 −2394−1016 1791 A C ATOM 1519 OG SER A 237 26.175 21.121 33.011 1.00 66.90 AO ANISOU 1519 OG SER A 237 7179 11836 6404 −2452 −983 1552 A O ATOM 1520N LEU A 238 28.921 21.050 35.659 1.00 54.51 A N ANISOU 1520 N LEU A 2385837 9400 5475 −2375 −1070 1741 A N ATOM 1521 CA LEU A 238 29.810 19.97636.056 1.00 47.92 A C ANISOU 1521 CA LEU A 238 5070 8408 4731 −2406−1054 1495 A C ATOM 1522 C LEU A 238 29.481 18.687 35.297 1.00 49.37 A CANISOU 1522 C LEU A 238 5305 8785 4667 −2489 −972 1118 A C ATOM 1523 OLEU A 238 28.369 18.154 35.444 1.00 53.99 A O ANISOU 1523 O LEU A 2385959 9366 5188 −2596 −1012 924 A O ATOM 1524 CB LEU A 238 29.710 19.76237.563 1.00 44.25 A C ANISOU 1524 CB LEU A 238 4802 7429 4583 −2342−1092 1438 A C ATOM 1525 CG LEU A 238 30.839 18.912 38.175 1.00 59.77 AC ANISOU 1525 CG LEU A 238 6878 9110 6723 −2275 −1028 1251 A C ATOM 1526CD1 LEU A 238 32.191 19.599 38.034 1.00 61.15 A C ANISOU 1526 CD1 LEU A238 6929 9304 6999 −2152 −983 1444 A C ATOM 1527 CD2 LEU A 238 30.57118.586 39.640 1.00 58.93 A C ANISOU 1527 CD2 LEU A 238 6966 8542 6884−2240 −1085 1179 A C ATOM 1528 N PRO A 239 30.414 18.148 34.510 1.0048.71 A N ANISOU 1528 N PRO A 239 5186 8866 4456 −2450 −851 987 A N ATOM1529 CA PRO A 239 30.156 16.871 33.835 1.00 41.94 A C ANISOU 1529 CA PROA 239 4380 8168 3387 −2528 −763 603 A C ATOM 1530 C PRO A 239 29.86715.740 34.807 1.00 41.46 A C ANISOU 1530 C PRO A 239 4545 7709 3499−2564 −735 295 A C ATOM 1531 O PRO A 239 30.485 15.632 35.868 1.00 39.14A O ANISOU 1531 O PRO A 239 4386 7011 3474 −2471 −733 321 A O ATOM 1532CB PRO A 239 31.456 16.604 33.066 1.00 45.72 A C ANISOU 1532 CB PRO A239 4792 8802 3777 −2448 −639 557 A C ATOM 1533 CG PRO A 239 32.48317.486 33.730 1.00 46.05 A C ANISOU 1533 CG PRO A 239 4808 8613 4075−2323 −653 845 A C ATOM 1534 CD PRO A 239 31.725 18.707 34.135 1.0045.26 A C ANISOU 1534 CD PRO A 239 4643 8499 4053 −2342 −780 1174 A CATOM 1535 N LYS A 240 28.918 14.885 34.415 1.00 44.16 A N ANISOU 1535 NLYS A 240 4926 8178 3675 −2695 −703 −7 A N ATOM 1536 CA LYS A 240 28.68013.597 35.068 1.00 48.55 A C ANISOU 1536 CA LYS A 240 5703 8401 4343−2747 −617 −362 A C ATOM 1537 C LYS A 240 29.616 12.581 34.428 1.0046.22 A C ANISOU 1537 C LYS A 240 5441 8167 3951 −2708 −466 −625 A CATOM 1538 O LYS A 240 29.315 12.018 33.374 1.00 56.94 A O ANISOU 1538 OLYS A 240 6715 9863 5058 −2799 −399 −856 A O ATOM 1539 CB LYS A 24027.218 13.168 34.939 1.00 53.94 A C ANISOU 1539 CB LYS A 240 6397 91884909 −2924 −633 −588 A C ATOM 1540 CG LYS A 240 26.191 14.176 35.4601.00 52.00 A C ANISOU 1540 CG LYS A 240 6116 8871 4771 −2881 −738 −320 AC ATOM 1541 CD LYS A 240 26.283 14.383 36.947 1.00 54.48 A C ANISOU 1541CD LYS A 240 6611 8681 5409 −2804 −781 −188 A C ATOM 1542 CE LYS A 24025.614 15.680 37.378 1.00 54.97 A C ANISOU 1542 CE LYS A 240 6609 87075570 −2699 −885 154 A C ATOM 1543 NZ LYS A 240 25.958 15.990 38.818 1.0055.51 A N1+ ANISOU 1543 NZ LYS A 240 6832 8309 5950 −2571 −926 306 A N1+ATOM 1544 N LEU A 241 30.769 12.365 35.054 1.00 42.94 A N ANISOU 1544 NLEU A 241 5140 7445 3732 −2561 −418 −598 A N ATOM 1545 CA LEU A 24131.767 11.481 34.478 1.00 44.65 A C ANISOU 1545 CA LEU A 241 5380 77143872 −2500 −283 −825 A C ATOM 1546 C LEU A 241 31.309 10.031 34.511 1.0052.03 A C ANISOU 1546 C LEU A 241 6496 8496 4775 −2587 −155 −1247 A CATOM 1547 O LEU A 241 30.438 9.643 35.304 1.00 49.75 A O ANISOU 1547 OLEU A 241 6372 7926 4604 −2662 −154 −1356 A O ATOM 1548 CB LEU A 24133.075 11.602 35.233 1.00 43.41 A C ANISOU 1548 CB LEU A 241 5300 72523943 −2306 −272 −710 A C ATOM 1549 CG LEU A 241 33.764 12.954 35.2331.00 45.29 A C ANISOU 1549 CG LEU A 241 5360 7598 4251 −2211 −360 −330 AC ATOM 1550 CD1 LEU A 241 35.139 12.734 35.839 1.00 41.88 A C ANISOU1550 CD1 LEU A 241 4999 6897 4014 −2022 −316 −348 A C ATOM 1551 CD2 LEUA 241 33.847 13.528 33.814 1.00 50.48 A C ANISOU 1551 CD2 LEU A 241 57738776 4632 −2271 −343 −220 A C ATOM 1552 N ASP A 242 31.924 9.224 33.6381.00 49.18 A N ANISOU 1552 N ASP A 242 6107 8317 4262 −2580 −31 −1492 AN ATOM 1553 CA ASP A 242 31.718 7.780 33.640 1.00 61.67 A C ANISOU 1553CA ASP A 242 7869 9727 5835 −2644 123 −1909 A C ATOM 1554 C ASP A 24232.673 7.148 34.654 1.00 66.37 A C ANISOU 1554 C ASP A 242 8709 98366674 −2467 195 −1966 A C ATOM 1555 O ASP A 242 33.891 7.191 34.471 1.0074.39 A O ANISOU 1555 O ASP A 242 9683 10877 7705 −2314 217 −1913 A OATOM 1556 CB ASP A 242 31.924 7.194 32.241 1.00 68.49 A C ANISOU 1556 CBASP A 242 8585 11013 6423 −2712 223 −2160 A C ATOM 1557 CG ASP A 24231.648 5.684 32.177 1.00 82.35 A C ANISOU 1557 CG ASP A 242 10516 126038172 −2798 400 −2619 A C ATOM 1558 OD1 ASP A 242 30.621 5.232 32.7471.00 88.58 A O ANISOU 1558 OD1 ASP A 242 11444 13166 9046 −2919 433−2777 A O ATOM 1559 OD2 ASP A 242 32.442 4.948 31.536 1.00 86.95 A O1−ANISOU 1559 OD2 ASP A 242 11091 13281 8667 −2752 517 −2831 A O1− ATOM1560 N GLN A 243 32.119 6.561 35.724 1.00 61.50 A N ANISOU 1560 N GLN A243 8347 8781 6240 −2478 238 −2078 A N ATOM 1561 CA GLN A 243 32.8995.956 36.797 1.00 58.37 A C ANISOU 1561 CA GLN A 243 8213 7897 6066−2287 302 −2122 A C ATOM 1562 C GLN A 243 32.560 4.472 36.948 1.00 67.62A C ANISOU 1562 C GLN A 243 9641 8795 7256 −2343 499 −2514 A C ATOM 1563O GLN A 243 31.439 4.051 36.630 1.00 66.00 A O ANISOU 1563 O GLN A 2439430 8664 6982 −2531 562 −2688 A O ATOM 1564 CB GLN A 243 32.650 6.66838.136 1.00 49.50 A C ANISOU 1564 CB GLN A 243 7203 6431 5174 −2204 182−1852 A C ATOM 1565 CG GLN A 243 32.934 8.176 38.167 1.00 42.32 A CANISOU 1565 CG GLN A 243 6062 5720 4296 −2149 −4 −1456 A C ATOM 1566 CDGLN A 243 34.427 8.513 38.178 1.00 49.20 A C ANISOU 1566 CD GLN A 2436859 6603 5233 −1932 −26 −1328 A C ATOM 1567 OE1 GLN A 243 35.295 7.62838.170 1.00 51.33 A O ANISOU 1567 OE1 GLN A 243 7245 6742 5515 −1803 83−1530 A O ATOM 1568 NE2 GLN A 243 34.728 9.807 38.190 1.00 43.34 A NANISOU 1568 NE2 GLN A 243 5915 6017 4534 −1892 −159 −1000 A N ATOM 1569N PRO A 244 33.512 3.650 37.422 1.00 70.28 A N ANISOU 1569 N PRO A 24410189 8812 7701 −2152 606 −2641 A N ATOM 1570 CA PRO A 244 33.291 2.20637.623 1.00 75.79 A C ANISOU 1570 CA PRO A 244 11103 9214 8480 −2098 794−2887 A C ATOM 1571 C PRO A 244 32.114 1.882 38.541 1.00 74.33 A CANISOU 1571 C PRO A 244 11039 8727 8477 −2086 807 −2776 A C ATOM 1572 OPRO A 244 32.092 2.435 39.640 1.00 74.30 A O ANISOU 1572 O PRO A 24411138 8467 8624 −1979 712 −2559 A O ATOM 1573 CB PRO A 244 34.603 1.73838.252 1.00 73.27 A C ANISOU 1573 CB PRO A 244 10976 8587 8277 −1819 838−2904 A C ATOM 1574 CG PRO A 244 35.623 2.703 37.720 1.00 68.87 A CANISOU 1574 CG PRO A 244 10197 8350 7622 −1766 721 −2771 A C ATOM 1575CD PRO A 244 34.922 4.025 37.652 1.00 63.85 A C ANISOU 1575 CD PRO A 2449337 7961 6960 −1897 544 −2470 A C HETATM 1576 C1 NAG A 1076 47.8496.531 51.063 1.00 44.10 A C HETATM 1577 C2 NAG A 1076 47.870 5.04350.676 1.00 57.73 A C HETATM 1578 C3 NAG A 1076 48.738 4.258 51.649 1.0062.74 A C HETATM 1579 C4 NAG A 1076 50.134 4.860 51.736 1.00 73.55 A CHETATM 1580 C5 NAG A 1076 50.074 6.367 52.006 1.00 66.00 A C HETATM 1581C6 NAG A 1076 51.433 7.025 51.850 1.00 77.39 A C HETATM 1582 C7 NAG A1076 45.835 4.269 49.503 1.00 69.45 A C HETATM 1583 C8 NAG A 1076 44.4863.638 49.680 1.00 73.06 A C HETATM 1584 N2 NAG A 1076 46.531 4.47250.630 1.00 67.87 A N HETATM 1585 O3 NAG A 1076 48.812 2.907 51.207 1.0061.48 A O HETATM 1586 O4 NAG A 1076 50.834 4.262 52.824 1.00 86.01 A OHETATM 1587 O5 NAG A 1076 49.177 7.032 51.099 1.00 44.53 A O HETATM 1588O6 NAG A 1076 51.359 8.424 51.609 1.00 83.18 A O HETATM 1589 O7 NAG A1076 46.264 4.592 48.392 1.00 59.22 A O HETATM 1590 C1 NAG A 2076 51.7043.148 52.511 1.00 89.75 A C HETATM 1591 C2 NAG A 2076 52.777 3.07653.594 1.00 84.61 A C HETATM 1592 C3 NAG A 2076 53.721 1.911 53.318 1.0085.99 A C HETATM 1593 C4 NAG A 2076 52.918 0.618 53.266 1.00 85.51 A CHETATM 1594 C5 NAG A 2076 51.825 0.728 52.199 1.00 92.45 A C HETATM 1595C6 NAG A 2076 50.898 −0.470 52.172 1.00 89.30 A C HETATM 1596 C7 NAG A2076 53.343 5.169 54.748 1.00 82.17 A C HETATM 1597 C8 NAG A 2076 54.1446.438 54.698 1.00 79.48 A C HETATM 1598 N2 NAG A 2076 53.500 4.33453.709 1.00 82.34 A N HETATM 1599 O3 NAG A 2076 54.714 1.836 54.336 1.0095.43 A O HETATM 1600 O4 NAG A 2076 53.761 −0.499 53.000 1.00 83.01 A OHETATM 1601 O5 NAG A 2076 50.999 1.883 52.440 1.00 95.99 A O HETATM 1602O6 NAG A 2076 49.529 −0.093 52.076 1.00 83.95 A O HETATM 1603 O7 NAG A2076 52.591 4.908 55.693 1.00 78.70 A O HETATM 1604 C1 NAG A 1123 47.70331.112 51.348 1.00 63.05 A C HETATM 1605 C2 NAG A 1123 48.304 32.50051.518 1.00 72.32 A C HETATM 1606 C3 NAG A 1123 48.864 32.659 52.9301.00 76.90 A C HETATM 1607 C4 NAG A 1123 47.778 32.420 53.977 1.00 85.17A C HETATM 1608 C5 NAG A 1123 47.071 31.072 53.759 1.00 75.98 A C HETATM1609 C6 NAG A 1123 45.790 30.947 54.564 1.00 66.09 A C HETATM 1610 C7NAG A 1123 49.136 33.360 49.363 1.00 71.99 A C HETATM 1611 C8 NAG A 112350.348 33.540 48.494 1.00 68.30 A C HETATM 1612 N2 NAG A 1123 49.34432.750 50.534 1.00 74.04 A N HETATM 1613 O3 NAG A 1123 49.408 33.96953.060 1.00 73.86 A O HETATM 1614 O4 NAG A 1123 48.360 32.443 55.2831.00 97.38 A O HETATM 1615 O5 NAG A 1123 46.695 30.884 52.381 1.00 77.46A O HETATM 1616 O6 NAG A 1123 45.575 29.627 55.047 1.00 67.23 A O HETATM1617 O7 NAG A 1123 48.021 33.741 49.014 1.00 73.84 A O HETATM 1618 C1NAG A 2123 47.809 33.471 56.171 1.00 101.53 A C HETATM 1619 C2 NAG A2123 48.615 33.558 57.507 1.00 98.63 A C HETATM 1620 C3 NAG A 212348.119 34.727 58.370 1.00 103.69 A C HETATM 1621 C4 NAG A 2123 48.08336.025 57.574 1.00 109.93 A C HETATM 1622 C5 NAG A 2123 47.220 35.82256.331 1.00 112.93 A C HETATM 1623 C6 NAG A 2123 47.087 37.055 55.4601.00 112.42 A C HETATM 1624 C7 NAG A 2123 49.181 31.201 57.925 1.0093.84 A C HETATM 1625 C8 NAG A 2123 48.955 30.016 58.818 1.00 90.46 A CHETATM 1626 N2 NAG A 2123 48.513 32.311 58.250 1.00 93.20 A N HETATM1627 O3 NAG A 2123 48.935 34.886 59.527 1.00 100.98 A O HETATM 1628 O4NAG A 2123 47.558 37.076 58.380 1.00 108.32 A O HETATM 1629 O5 NAG A2123 47.794 34.783 55.522 1.00 111.35 A O HETATM 1630 O6 NAG A 212348.292 37.378 54.780 1.00 114.76 A O HETATM 1631 O7 NAG A 2123 49.93231.153 56.951 1.00 98.48 A O HETATM 1632 C1 NAG A 1144 54.671 15.65635.163 1.00 37.73 A C HETATM 1633 C2 NAG A 1144 54.274 14.196 35.0651.00 37.73 A C HETATM 1634 C3 NAG A 1144 55.503 13.302 34.960 1.00 50.44A C HETATM 1635 C4 NAG A 1144 56.531 13.777 33.938 1.00 51.62 A C HETATM1636 C5 NAG A 1144 56.778 15.283 34.077 1.00 47.74 A C HETATM 1637 C6NAG A 1144 57.543 15.877 32.914 1.00 55.32 A C HETATM 1638 C7 NAG A 114452.472 12.987 36.222 1.00 51.27 A C HETATM 1639 C8 NAG A 1144 51.85512.689 37.554 1.00 50.70 A C HETATM 1640 N2 NAG A 1144 53.515 13.81436.241 1.00 42.75 A N HETATM 1641 O3 NAG A 1144 55.056 11.989 34.6461.00 60.19 A O HETATM 1642 O4 NAG A 1144 57.685 12.994 34.250 1.00 60.95A O HETATM 1643 O5 NAG A 1144 55.528 15.984 34.100 1.00 47.14 A O HETATM1644 O6 NAG A 1144 56.804 16.883 32.230 1.00 61.36 A O HETATM 1645 O7NAG A 1144 52.016 12.538 35.177 1.00 59.72 A O HETATM 1646 C1 NAG A 214458.955 12.890 33.530 1.00 83.73 A C HETATM 1647 C2 NAG A 2144 59.53911.488 33.820 1.00 94.96 A C HETATM 1648 C3 NAG A 2144 60.887 11.31533.114 1.00 103.58 A C HETATM 1649 C4 NAG A 2144 60.782 11.691 31.6391.00 103.06 A C HETATM 1650 C5 NAG A 2144 60.091 13.044 31.444 1.0093.13 A C HETATM 1651 C6 NAG A 2144 59.775 13.317 29.996 1.00 88.48 A CHETATM 1652 C7 NAG A 2144 58.862 10.457 35.937 1.00 89.36 A C HETATM1653 C8 NAG A 2144 59.149 10.326 37.405 1.00 83.94 A C HETATM 1654 N2NAG A 2144 59.684 11.256 35.247 1.00 91.04 A N HETATM 1655 O3 NAG A 214461.315 9.960 33.242 1.00 102.99 A O HETATM 1656 O4 NAG A 2144 62.07811.733 31.048 1.00 106.07 A O HETATM 1657 O5 NAG A 2144 58.829 13.06632.129 1.00 93.17 A O HETATM 1658 O6 NAG A 2144 59.024 12.239 29.4531.00 86.36 A O HETATM 1659 O7 NAG A 2144 57.921 9.870 35.399 1.00 93.33A O HETATM 1660 C1 NAG A 1228 54.131 12.661 30.331 1.00 73.34 A C HETATM1661 C2 NAG A 1228 55.401 12.376 29.460 1.00 83.15 A C HETATM 1662 C3NAG A 1228 55.578 10.871 29.213 1.00 92.37 A C HETATM 1663 C4 NAG A 122855.493 10.061 30.498 1.00 93.75 A C HETATM 1664 C5 NAG A 1228 54.19410.379 31.232 1.00 90.93 A C HETATM 1665 C6 NAG A 1228 54.052 9.63132.538 1.00 89.70 A C HETATM 1666 C7 NAG A 1228 55.953 14.259 27.9621.00 62.50 A C HETATM 1667 C8 NAG A 1228 55.801 14.820 26.580 1.00 61.44A C HETATM 1668 N2 NAG A 1228 55.354 13.082 28.186 1.00 77.40 A N HETATM1669 O3 NAG A 1228 56.846 10.634 28.609 1.00 97.10 A O HETATM 1670 O4NAG A 1228 55.536 8.679 30.152 1.00 98.74 A O HETATM 1671 O5 NAG A 122854.141 11.786 31.533 1.00 87.29 A O HETATM 1672 O6 NAG A 1228 55.1549.888 33.397 1.00 97.52 A O HETATM 1673 O7 NAG A 1228 56.570 14.85528.842 1.00 51.25 A O HETATM 1674 C1 NAG A 2228 56.712 7.979 30.648 1.00105.75 A C HETATM 1675 O2 NAG A 2228 56.432 6.506 30.363 1.00 109.04 A CHETATM 1676 O3 NAG A 2228 57.551 5.638 30.941 1.00 114.26 A C HETATM1677 O4 NAG A 2228 58.904 6.090 30.393 1.00 114.48 A C HETATM 1678 O5NAG A 2228 59.110 7.592 30.626 1.00 116.17 A C HETATM 1679 O6 NAG A 222860.393 8.132 30.026 1.00 116.84 A C HETATM 1680 O7 NAG A 2228 54.0725.897 30.099 1.00 103.32 A C HETATM 1681 O8 NAG A 2228 54.306 6.05828.626 1.00 104.80 A C HETATM 1682 N2 NAG A 2228 55.133 6.111 30.8861.00 105.10 A N HETATM 1683 O3 NAG A 2228 57.316 4.272 30.614 1.00114.03 A O HETATM 1684 O4 NAG A 2228 59.957 5.358 31.014 1.00 109.16 A OHETATM 1685 O5 NAG A 2228 58.018 8.350 30.070 1.00 113.09 A O HETATM1686 O6 NAG A 2228 60.410 8.042 28.608 1.00 118.16 A O HETATM 1687 O7NAG A 2228 52.975 5.578 30.552 1.00 99.40 A O TER 1688 NAG A 2228 HETATM1688 O HOH S 1 31.421 19.873 44.880 1.00 42.21 S O HETATM 1689 O HOH S 341.885 20.609 55.460 1.00 40.39 S O HETATM 1690 O HOH S 4 60.490 20.80851.222 1.00 39.49 S O HETATM 1691 O HOH S 5 29.548 12.649 29.611 1.0024.15 S O TER 1693 HOH S 5 END

1. A method for selection of a target plant LysM receptor for modifyingthe target plant LysM receptor to have a desired receptorcharacteristic, comprising: (a) providing a structural model, amolecular model, a surface characteristics model, and/or anelectrostatic potential model of a donor plant LysM receptor having thedesired receptor characteristic and two or more potential target plantLysM receptors; (b) comparing each of the two or more potential targetplant LysM receptors with the structural model, the molecular model, thesurface characteristics model, and/or the electrostatic potential modelof the donor plant LysM receptor, and/or comparing each of the two ormore potential target plant LysM receptors with the donor plant LysMreceptor using structural overlay; and (c) selecting the potentialtarget plant LysM receptor with a suitable match for the donor plantLysM receptor to be the target plant LysM receptor.
 2. The method ofclaim 1, wherein the criteria for determining that the potential targetplant LysM receptor is a suitable match for the donor plant LysMreceptor in step (c) are selected from the group consisting of goodnessof fit to template structure; similarity; phylogenetic relation; surfacepotential; coverage to template structure; GMQE, QMEAN, and LocalQuality estimates from SWISS-Model; and any combination thereof.
 3. Themethod of claim 1, wherein the structural model of the donor plant LysMreceptor is a protein crystal structure, a molecular model, a cryo-EMstructure, and a NMR structure.
 4. The method of claim 1, wherein thedonor plant LysM receptor model is of an entire ectodomain and the twoor more potential target plant LysM receptor models are of entireectodomains.
 5. The method of claim 1, wherein the donor plant LysMreceptor model is of a LysM1 domain, a LysM2 domain, a LysM3 domain, orany combination thereof and the two or more potential target plant LysMreceptor models are of LysM1 domains, LysM2 domains, LysM3 domains, orany combination thereof.
 6. The method of claim 1, wherein the donorplant LysM receptor is Medicago NFP, Medicago LYK3, Lotus NFR1, LotusNFR5, Lotus LYS11, or Arabidopsis CERK1.
 7. The method of claim 6,wherein the two or more target plant LysM receptors are additionallycompared to Lotus CERK6.
 8. The method of claim 1, wherein the two ormore potential target plant LysM receptor polypeptides are all from thesame plant species or plant variety.
 9. The method of claim 1, whereinthe desired receptor characteristic is affinity, selectivity, and/orspecificity for an oligosaccharide or class of oligosaccharides.
 10. Themethod of claim 1, wherein the desired receptor characteristic isbinding kinetics for an oligosaccharide or class of oligosaccharides,wherein the binding kinetics comprise off-rate and on-rate.
 11. Themethod of claim 9, wherein the class of oligosaccharides is selectedfrom the group consisting of LCOs, COs, beta-glucans,cyclic-beta-glucans, exopolysaccharides, and optionally LPS.
 12. Themethod of claim 11, wherein the class of oligosaccharides is LCOs orCOs.
 13. The method of claim 12, wherein the class of oligosaccharidesis LCOs, optionally produced by a produced by a nitrogen-fixing bacteriaoptionally selected from the group consisting of Mesorhizobium loti,Mesorhizobium huakuii, Mesorhizobium mediterraneum, Mesorhizobiumciceri, Mesorhizobium spp., Rhizobium mongolense, Rhizobium tropici,Rhizobium etli phaseoli, Rhizobium giardinii, Rhizobium leguminosarumoptionally R. leguminosarum trifolii, R. leguminosarum viciae, and R.leguminosarum phaseoli, Burkholderiales optionally symbionts of Mimosa,Sinorhizobium meliloti, Sinorhizobium medicae, Sinorhizobium fredii,Sinorhizobium NGR234, Azorhizobium caulinodans, Bradyrhizobiumjaponicum, Bradyrhizobium elkanii, Bradyrhizobium liaonginense, Frankiaspp., and any combination thereof; or optionally produced by amycorrhizal fungi optionally selected from the group consisting ofAcaulosporaceae spp., Diversisporaceae spp., Gigasporaceae spp.,Pacisporaceae spp., Funneliformis spp., Glomus spp., Rhizophagus spp.,Sclerocystis spp., Septoglomus spp., Claroideoglomus spp., Ambisporaspp., Archaeospora spp., Geosiphon pyriformis, Paraglomus spp., otherspecies in the division Glomeromycota, and any combination thereof. 14.The method of claim 13, wherein at least one of the LCOs is M. loti LCO,S. meliloti LCO-IV, or S. meliloti LCO-V.
 15. The method of claim 1,further comprising step (d): identifying one or more amino acid residuesfor modification in the target LysM receptor by comparing amino acidresidues of a first oligosaccharide binding feature in the donor plantLysM receptor with the corresponding amino acid residues in the targetplant LysM receptor, and optionally identifying one or more amino acidresidues for modification in the target LysM receptor by comparing aminoacid residues of a second oligosaccharide binding feature in the donorplant LysM receptor with the corresponding amino acid residues in thetarget plant LysM receptor.
 16. The method of claim 15, furthercomprising step (e): generating a modified plant LysM receptor whereinthe one or more amino acid residues in the first oligosaccharide bindingfeature of the target plant LysM receptor have been substituted withcorresponding amino acid residues from the donor plant LysM receptor;generating a modified plant LysM receptor wherein the one or more aminoacid residues in the second oligosaccharide binding feature of thetarget plant LysM receptor have been substituted with correspondingamino acid residues from the donor plant LysM receptor; or generating amodified plant LysM receptor wherein the one or more amino acid residuesin the first oligosaccharide binding feature of the target plant LysMreceptor have been substituted with corresponding amino acid residuesfrom the donor plant LysM receptor and the one or more amino acidresidues in the second oligosaccharide binding feature of the targetplant LysM receptor have been substituted with corresponding amino acidresidues from the donor plant LysM receptor.
 17. The method of claim 16,wherein the first oligosaccharide binding feature is a hydrophobic patchon the surface of the LysM2 domain.
 18. The method of claim 16, whereinthe second oligosaccharide binding feature is a part of the LysM1 domainof the donor plant LysM receptor.
 19. A modified plant LysM receptorproduced using the method of claim 1, wherein the modified plant LysMreceptor comprises a LysM2 domain modified to comprise a hydrophobicpatch on the surface of the LysM2 domain.
 20. A modified plant LysMreceptor produced using the method of claim 1, wherein the modifiedplant LysM receptor comprises a first LysM1 domain modified to replaceat least part of the first LysM1 domain with at least part of a secondLysM1 domain.